Lipid analogs and liposomes comprising same

ABSTRACT

A polymeric compound is disclosed herein, having the general formula I: 
                         
wherein m, n, X, Y, Z and L are as defined herein. Further disclosed herein are lipid bilayers comprising at least one bilayer-forming lipid and the aforementioned polymeric compound, and liposomes comprising such a bilayer, as well as methods, uses and compositions utilizing such bilayers and/or liposomes for reducing a friction coefficient of a surface and/or for inhibiting biofilm formation.

RELATED APPLICATIONS

This application is a National Phase of PCT Patent Application No.PCT/IL2016/051372 having International filing date of Dec. 22, 2016,which claims the benefit of priority of Israel Patent Application No.243285 filed on Dec. 22, 2015. The contents of the above applicationsare all incorporated by reference as if fully set forth herein in theirentirety.

SEQUENCE LISTING STATEMENT

The ASCII file, entitled 74172SequenceListing.txt, created on Jun. 21,2018, comprising 734 bytes, submitted concurrently with the filing ofthis application is incorporated herein by reference. The sequencelisting submitted herewith is identical to the sequence listing formingpart of the international application.

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to materialscience and, more particularly, but not exclusively, to novel polymericcompounds usable, inter alia, for forming liposomes.

Phosphatidylcholine (PC) liposomes at surfaces are known to be extremelygood lubricants, even at high pressures [Goldberg et al., Adv Materials2011, 23:3517-3521; Goldberg et al., Biophys J 2011, 100:2403-2411;Sorkin et al., Biomaterials 2014, 34:5465-5475].

Aggregation of liposomes into macroscopic aggregates can interfere withthe use of liposomes in different ways. Large aggregates can precipitateand sediment out of a dispersion, rendering the dispersion unusable;aggregates larger than about 200-300 nm scatter visible light, leadingto turbidity, which may interfere with a use of liposomes in whichtransparency is important; and furthermore, large aggregates injectedinto the body are more prone to protein adsorption, and to attack andremoval by macrophages [Moghimi & Szebeni, Prog Lipid Res 2003,42:463-478].

PEGylated PC small unilamellar vesicles (SUVs) have been used for drugdelivery, wherein PEG brushes are incorporated in the membrane bilayer;these brushes extend out from the SUV surfaces and sterically-stabilizethem against aggregation [Harris & Chess, Nat Rev Drug Discov 2003,2:214-221]. However, PEGylation was reported to reduce the efficiency ofSUVs for lubrication purposes at high pressures (such as in joints), asthe PEG chains are not highly hydrated and do not in themselves formgood lubricants at high pressures [Goldberg et al., Adv Materials 2011,23:3517-3521].

U.S. Pat. No. 8,617,592 describes block copolymers and conjugatescomprising a zwitterionic poly(carboxybetaine), poly(sulfobetaine) orpoly(phosphobetaine) block, and a hydrophobic block, which self-assembleinto particles, and the use of such particles for delivering therapeuticand diagnostic agents.

Chen et al. [Science 2009, 323:1698-1702] describes effectivelubrication by poly[2-(methacryloyloxy)ethyl phosphorylcholine] (PMPC)brushes, and attributes this phenomenon to strong hydration of thezwitterionic monomers.

Additional background art includes Goldberg & Klein [Chem Phys Lipids2012, 165:374-381]; International Patent Application Nos.PCT/IL2014/050604 (published as WO 2015/001564), PCT/IL2015/050605 andPCT/IL2015/050606; and Israel Patent Application No. 234929.

SUMMARY OF THE INVENTION

According to an aspect of some embodiments of the invention, there isprovided a polymeric compound having the general formula I:

wherein:m is zero or a positive integer;n is an integer which is at least 1, wherein when X does not comprise aphosphate group, n is at least 2;X is a lipid moiety;Y is a backbone unit which forms a polymeric backbone;L is absent or is a linking moiety; andZ has the general formula II:

wherein:A is a substituted or unsubstituted hydrocarbon;B is an oxygen atom or is absent; andR₁-R₃ are each independently selected from the group consisting ofhydrogen, alkyl, cycloalkyl, heteroalicyclic, aryl and heteroaryl.

According to an aspect of some embodiments of the invention, there isprovided a lipid bilayer comprising at least one bilayer-forming lipidand the polymeric compound according to any of the respectiveembodiments described herein.

According to an aspect of some embodiments of the invention, there isprovided a liposome comprising at least one lipid bilayer according toany of the respective embodiments described herein.

According to an aspect of some embodiments of the invention, there isprovided a lubricant composition comprising liposomes according to anyof the respective embodiments described herein and a carrier.

According to an aspect of some embodiments of the invention, there isprovided a method of reducing a friction coefficient of a surface, themethod comprising contacting the surface with liposomes according to anyof the respective embodiments described herein.

According to an aspect of some embodiments of the invention, there isprovided a use of a liposome of according to any of the respectiveembodiments described herein in the manufacture of a medicament fortreating a synovial joint disorder associated with an increased frictioncoefficient of an articular surface in the synovial joint.

According to an aspect of some embodiments of the invention, there isprovided a method of inhibiting biofilm formation on a surface of asubstrate, the method comprising contacting the substrate with acomposition which comprises liposomes to any of the respectiveembodiments described herein.

According to an aspect of some embodiments of the invention, there isprovided an article of manufacture comprising a composition-of-matter,the composition-of-matter comprising a substrate coated, on at least aportion of a surface thereof, by a lipid bilayer according to any of therespective embodiments described herein.

According to some embodiments of the invention, Y is a substituted orunsubstituted alkylene unit.

According to some embodiments of the invention, Y is a substituted orunsubstituted ethylene unit.

According to some embodiments of the invention, Y has the formula—CR₄R₅—CR₆D-, wherein:

when Y is a backbone unit which is not attached to L or Z, D is R₇; andwhen Y is a backbone unit which is attached to L or Z, D is a covalentbond or a linking group attaching Y to L or Z, the linking group beingselected from the group consisting of —O—, —S—, alkylene, arylene,sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl,thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl,N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, andamino; and

R₄-R₇ are each independently selected from the group consisting ofhydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl,heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy,thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, azide, azo,phosphate, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea,thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, and amino.

According to some embodiments of the invention, R₄ and R₅ are eachhydrogen.

According to some embodiments of the invention, R₆ is hydrogen.

According to some embodiments of the invention, the linking group isselected from the group consisting of —O—, —C(═O)O—, —C(═O)NH— andphenylene.

According to some embodiments of the invention, the linking group is—C(═O)O—.

According to some embodiments of the invention, L is a substituted orunsubstituted hydrocarbon from 1 to 10 carbon atoms in length.

According to some embodiments of the invention, L is a substituted orunsubstituted ethylene group.

According to some embodiments of the invention, B is an oxygen atom.

According to some embodiments of the invention, A is a substituted orunsubstituted hydrocarbon from 1 to 4 carbon atoms in length.

According to some embodiments of the invention, A is a substituted orunsubstituted ethylene group.

According to some embodiments of the invention, R₁-R₃ are eachindependently hydrogen or C₁₋₄-alkyl.

According to some embodiments of the invention, R₁-R₃ are each methyl.

According to some embodiments of the invention, n is at least 3.

According to some embodiments of the invention, n is in a range of from3 to 1,000, and m is in a range of from 0 to 1,000.

According to some embodiments of the invention, n is in a range of from5 to 50, and m is in a range of from 0 to 50.

According to some embodiments of the invention, at least a portion of Y,L and/or Z comprise at least one targeting moiety.

According to some embodiments of the invention, the polymeric compoundhas the general formula Ib:

wherein:

T is a unit of Y which comprises at least one targeting moiety, asdescribed hereinabove;

X and T are attached to distal termini of the polymeric compound; and

X, Y, L, Z, n and m are as defined for general formula I, with theproviso that m is a positive integer.

According to some embodiments of the invention, the lipid is selectedfrom the group consisting of a fatty acid, a monoglyceride, adiglyceride, a triglyceride, a glycerophospholipid, a sphingolipid, anda sterol.

According to some embodiments of the invention, the glycerophospholipidis selected from the group consisting of a phosphatidyl ethanolamine, aphosphatidyl serine, a phosphatidyl glycerol and a phosphatidylinositol.

According to some embodiments of the invention, X has the generalformula III:

wherein:

W₁ and W₂ are each independently selected from the group consisting ofhydrogen, alkyl, alkenyl, alkynyl and acyl, wherein at least one of W₁and W₂ is not hydrogen;

J is —P(═O)(OH)—O— or absent;

K is a substituted or unsubstituted hydrocarbon from 1 to 10 carbonatoms in length;

M is a linking group selected from the group consisting of —O—, —S—,amino, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl,thiocarbonyl, urea, thiourea, carbamyl, thiocarbamyl, amido, carboxy,and sulfonamide, or absent; and

Q is a substituted or unsubstituted hydrocarbon from 1 to 10 carbonatoms in length, or absent,

wherein when M is absent, Q is also absent.

According to some embodiments of the invention, J is —P(═O)(OH)—O— and Kis selected from the group consisting of an ethanolamine moiety, aserine moiety, a glycerol moiety and an inositol moiety.

According to some embodiments of the invention, M is amido.

According to some embodiments of the invention, Q is a substituted orunsubstituted methylene group.

According to some embodiments of the invention, Q is dimethylmethylene(—C(CH₃)₂—).

According to some embodiments of the invention, J, M and Q are eachabsent.

According to some embodiments of the invention, K is —C(═O)—C(CH₃)₂—.

According to some embodiments of the invention, at least one of W₁ andW₂ is alkyl, alkenyl, alkynyl or acyl, being from 10 to 30 carbon atomsin length.

According to some embodiments of the invention, the lipid moietycomprises at least one fatty acid moiety selected from the groupconsisting of lauroyl, myristoyl, palmitoyl, stearoyl, palmitoleoyl,oleoyl, and linoleoyl.

According to some embodiments of the invention, a molar ratio of thebilayer-forming lipid and the polymeric compound in a lipid bilayerdescribed herein is in a range of from 5:1 to 5,000:1.

According to some embodiments of the invention, a liposome describedherein further comprises at least one functional moiety or agent, boundto a surface of the liposome and/or within a lipid bilayer and/or coreof the liposome.

According to some embodiments of the invention, the functional moiety oragent is a therapeutically active agent or moiety thereof, a labelingmoiety or agent and/or a targeting moiety or agent.

According to some embodiments of the invention, a carrier as describedherein comprises an aqueous liquid.

According to some embodiments of the invention, a lubricant compositionas described herein further comprises a water-soluble polymer.

According to some embodiments of the invention, a lubricant compositionaccording to any of the respective embodiments described herein is forlubricating a physiological surface, wherein the carrier is aphysiologically acceptable carrier.

According to some embodiments of the invention, a method according toany of the respective embodiments described herein comprises contactinga surface with a composition comprising the liposomes and a carrier.

According to some embodiments of the invention, a method according toany of the respective embodiments described herein further comprisescontacting a surface with a water-soluble polymer.

According to some embodiments of the invention, a surface as describedherein is a hydrogel surface.

According to some embodiments of the invention, a surface as describedherein is a contact lens surface.

According to some embodiments of the invention, a surface as describedherein is a physiological surface, and a carrier described herein is aphysiologically acceptable carrier.

According to some embodiments of the invention, the surface is anarticular surface of a synovial joint.

According to some embodiments of the invention, a liposome, method,medicament and/or composition according to any of the respectiveembodiments described herein is for use in the treatment of a synovialjoint disorder associated with an increased friction coefficient of anarticular surface in the synovial joint.

According to some embodiments of the invention, the synovial jointdisorder is selected from the group consisting of arthritis, traumaticjoint injury, locked joint, and joint injury associated with surgery.

Unless otherwise defined, all technical and/or scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which the invention pertains. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of embodiments of the invention, exemplarymethods and/or materials are described below. In case of conflict, thepatent specification, including definitions, will control. In addition,the materials, methods, and examples are illustrative only and are notintended to be necessarily limiting.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

Some embodiments of the invention are herein described, by way ofexample only, with reference to the accompanying drawings. With specificreference now to the drawings in detail, it is stressed that theparticulars shown are by way of example and for purposes of illustrativediscussion of embodiments of the invention. In this regard, thedescription taken with the drawings makes apparent to those skilled inthe art how embodiments of the invention may be practiced.

In the drawings:

FIG. 1 presents a ¹H-NMR spectrum and corresponding structure ofDSPE-PMPC(distearoylphosphatidylethanolamine-poly((O-(2-methacryloyloxyethyl)phosphorylcholine)) prepared according to exemplary embodiments of theinvention;

FIG. 2 is a graph showing distribution of liposome particle size byvolume following storage of HSPC (hydrogenated soy phosphatidylcholine)liposomes (red/dark line) and HSPC liposomes with DSPE-PMPC (green/lightline) for one month, as determined by dynamic light scatteringmeasurements;

FIG. 3 is an atomic force microscopy (AFM) image of a mica surface afterexposure to DSPE-PMPC-stabilized HSPC liposomes in water;

FIG. 4 is an AFM image of a mica surface after exposure toDSPE-PMPC-stabilized HSPC liposomes in water at pH 5 (after which the pH5 water was replaced by conductivity water (pH 5.8));

FIG. 5A is a graph showing the normal force (normalized as Fn/R in theDerjaguin approximation) as a function of distance (D) between two micasurfaces coated with DSPE-PMPC-stabilized HSPC (PMPC) orDSPE-PEG-stabilized HSPC (PEG) liposomes in conductivity water (filledsymbols indicate first approach at a given contact point, correspondingsymbols indicate second approach at the same contact point);

FIG. 5B is a graph showing the friction force (Fs) between two micasurfaces coated with DSPE-PMPC-stabilized HSPC liposomes (PMPC) orDSPE-PEG-stabilized HSPC liposomes (PEG) in conductivity water as afunction of applied loads (normal force Fn) (filled symbols indicatefirst approach at a given contact point, corresponding symbols indicatesecond approach at the same contact point; friction coefficients μ(defined as Fs/Fn) and pressures (P) corresponding to the highest loadsattained in a data set are indicated at the end points of the datasets);

FIG. 6A is a graph showing the normal force (normalized as Fn/R in theDerjaguin approximation) as a function of distance (D) between two micasurfaces coated with DSPE-PMPC-stabilized HSPC (PMPC) orDSPE-PEG-stabilized HSPC (PEG) liposomes in aqueous 0.15 M NaNO₃ (filledsymbols indicate first approach at a given contact point, correspondingsymbols indicate second approach at the same contact point);

FIG. 6B is a graph showing the friction force (Fs) between two micasurfaces coated with DSPE-PMPC-stabilized HSPC (PMPC) orDSPE-PEG-stabilized HSPC (PEG) liposomes in aqueous 0.15 M NaNO₃ as afunction of applied loads (normal force Fn) (filled symbols indicatefirst approach at a given contact point, corresponding symbols indicatesecond approach at the same contact point; friction coefficients μ(defined as Fs/Fn) and pressures (P) corresponding to the highest loadsattained in a data set are indicated at the end points of the datasets);

FIG. 7 is a is a graph showing distribution of micelle size by volumefollowing incubation of 20 mg/ml DSPE-PMPC in water for one day, asdetermined by dynamic light scattering measurements;

FIGS. 8A and 8B present photographs of a cornea-mimicking lens holder(FIG. 8A) and the same holder with a soft contact lens mounted in place(FIG. 8B), used in some of the experiments employing a tribometer formeasuring friction coefficients of contact lenses described in theExamples section herein;

FIG. 9 is a graph showing the friction coefficient of etafilcon Acontact lenses under loads of 3 or 10 grams (corresponding to meanpressures of 0.1 and 0.16 atmospheres, respectively) followingincubation for 2 days in PBS with small unilamellar vesicles (SUV) ofHSPC and DSPE-PMPC (HSPC/pMPC), HSPC and DSPE-PEG (HSPC/PEG) or DMPC andDPPE-PEG (DMPC/PEG), or in PBS without liposomes (PBS);

FIG. 10 is a graph showing the friction coefficient of narafilcon Acontact lenses under loads of 3 or 10 grams (corresponding to meanpressures of 0.18 and 0.26 atmospheres, respectively) followingincubation for 2 days in PBS with small unilamellar vesicles (SUV) ofHSPC and DSPE-PMPC (HSPC/pMPC) or HSPC and DSPE-PEG (HSPC/PEG), or inPBS without liposomes (PBS);

FIG. 11 is a histogram showing the distribution of apparent zetapotential of small unilamellar vesicles (SUV) of HSPC and DSG-PMPC(1,2-distearoyl-sn-glycerol-poly((O-(2-methacryloyloxyethyl)phosphorylcholine)) prepared according to exemplary embodiments of theinvention;

FIG. 12 is an AFM image of a mica surface after exposure toDSG-PMPC-stabilized HSPC liposomes in water (pH 5.8); and

FIG. 13 is a graph showing the friction force (Fs) between two micasurfaces coated with DSG-PMPC-stabilized HSPC liposomes in water as afunction of applied loads (normal force Fn) (friction coefficients μ(defined as Fs/Fn) and pressures (P) corresponding to the highest loadsattained in a data set are indicated at the end points of the datasets).

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention, in some embodiments thereof, relates to materialscience and, more particularly, but not exclusively, to novel polymericcompounds usable, inter alia, for forming liposomes.

Before explaining at least one embodiment of the invention in detail, itis to be understood that the invention is not necessarily limited in itsapplication to the details set forth in the following description orexemplified by the Examples. The invention is capable of otherembodiments or of being practiced or carried out in various ways.

In a search for polymeric compounds capable of stabilizing lipid layerssuch as those of liposomes, the present inventors have designedcompounds comprising a lipid moiety and a polymeric moiety in whichmonomers include both phosphate and ammonium ionic groups, which aresimilar to phosphocholine and related head groups of liposome-formingphospholipids such as phosphatidylcholines.

While reducing the present invention to practice, the inventors haveuncovered that liposomes containing even small amounts of such alipid-containing polymeric compound exhibit considerably enhancedstability and are highly effective lubricants between sliding surfaces,being more effective lubricants than liposomes conjugated to PEG,particularly in a saline environment (e.g., a physiological environment)and/or at high pressures.

The inventors have further uncovered that such lipid-containingpolymeric compounds per se form stable micelles in an aqueousenvironment.

Embodiments of the present invention therefore relate to polymericcompounds which bear phosphocholine analogs as pendant groups and areconjugated to a lipid (e.g., phospholipid) moiety. Exemplary suchpolymeric compounds are represented by formula I. These polymericcompounds are also referred to herein as “lipid-containing polymericcompounds” or simply as “polymeric compounds”.

The lipid-containing polymeric compounds disclosed herein are capable ofstabilizing liposomes used for various applications (including in vivoapplications) against aggregation and fusion, thereby increasing shelflife, while retaining and even enhancing properties associated with thesurfaces of liposomes and other phospholipid layers, such asbiocompatibility, a high degree of hydration, and lubricant activity(e.g., by hydration lubrication). The disclosed polymeric compounds perse are also capable of forming stable micelles in an aqueous environmentwhich can be used as stable replacement for liposomes in variousapplications (including in vivo applications), such as lubrication,including lubrication of interfaces with physiological surfaces.

According to an aspect of some embodiments of the invention, there isprovided a polymeric compound having the general formula I:

wherein:

m is zero or a positive integer;

n is an integer which is at least 1;

X is a lipid moiety, wherein when X does not comprise a phosphate group,n is at least 2;

Y is a backbone unit which forms a polymeric backbone;

L is absent or is a linking moiety; and

Z has the general formula II:

wherein:

A is a substituted or unsubstituted hydrocarbon;

B is an oxygen atom or is absent; and

R₁-R₃ are each independently hydrogen, alkyl, cycloalkyl,heteroalicyclic, aryl or heteroaryl,

as described in more detail herein below.

Formula I may also be described herein simply as:X—[Y(-L-Z)]n[Y]m

which is to be regarded as interchangeable with the schematic depictionhereinabove.

Polymeric Moiety:

Herein, the term “polymeric” refers a compound having at least 2repeating units (and more preferably at least 3 repeating units), therepeating units being identical or similar. It is to be appreciated thatthe compound of general formula I is by definition polymeric when n isat least 2, as it comprises at least 2 of the backbone units representedby Y.

Herein, the phrase “polymeric moiety” refers to the portion of thepolymeric compound (according to any of the embodiments described hereinrelating to general formula I) which has the general formula Ia:

wherein m, n, Y, L and Z are as defined herein for general formula I.

Formula Ia may also be described herein simply as:[Y(-L-Z)]n[Y]m

which is to be regarded as interchangeable with the schematic depictionhereinabove.

Herein, the phrase “polymeric compound” further encompasses compoundshaving a “polymeric moiety” as described herein having one unit (e.g.,according to formula Ia wherein n is 1), provided that the lipid moietydescribed herein (e.g., the lipid moiety represented by X) has a similarunit. For example, when the lipid moiety comprises a phosphate group(e.g., the lipid moiety is a glycerophospholipid moiety), such that thelipid moiety has a phosphate group and a single unit of the polymericmoiety has a phosphate group, the two phosphate groups may be regardedas repeating units.

In preferred embodiments however, n is at least 2, such that thepolymeric moiety per se has at least two units. In some embodiments, nis at least 3.

As used herein, the term “backbone unit” refers to a repeating unit,wherein linkage of a plurality of the repeating unit (e.g., sequentiallinkage) forms a polymeric backbone. A plurality of linked repeatingunits per se is also referred to herein as a “polymeric backbone”.

As shown in formulas I and Ia, L and Z together form a pendant group ofat least a portion of the backbone units, which group is referred toherein for brevity simply as the “pendant group”.

Each backbone unit Y with pendant group (i.e., a unit represented byY(-L-Z), the number of which is represented by the variable n) and eachbackbone unit Y without a pendant group (the number of which isrepresented by the variable m) is also referred to herein as a“monomeric unit”.

A backbone unit may optionally be a residue of a polymerizable monomeror polymerizable moiety of a monomer. A wide variety of polymerizablemonomers and moieties will be known to the skilled person, and thestructure of the residues of such monomers which result uponpolymerization (e.g., monomeric units) will also be known to the skilledperson.

A “residue of a polymerizable monomer” refers to a modified form of apolymerizable monomer and/or a portion of a polymerizable monomer thatremains after polymerization.

A portion of a polymerizable monomer may be formed, for example, by acondensation reaction, e.g., wherein at least one atom or group (e.g., ahydrogen atom or hydroxyl group) in the monomer, and optionally at leasttwo atoms or groups (e.g., a hydrogen atom and a hydroxyl group) in themonomer, is replaced with a covalent bond with another polymerizablemonomer.

A modified form of a polymerizable monomer may be formed, for example,by ring-opening (wherein a covalent bond between two atoms in a ring isbroken, and the two atoms optionally each become linked to anotherpolymerizable monomer); and/or by adding to an unsaturated bond, whereinan unsaturated bond between two adjacent atoms is broken (e.g.,conversion of an unsaturated double bond to a saturated bond, orconversion of an unsaturated triple bond to an unsaturated double bond)and the two atoms optionally each become linked to another polymerizablemonomer.

A modified form of a polymerizable monomer may consist essentially ofthe same atoms as the original monomer, for example, different merely inthe rearrangement of covalent bonds, or alternatively, may have adifferent atomic composition, for example, wherein polymerizationincludes a condensation reaction (e.g., as described herein).

Examples of backbone units include, without limitation, substituted orunsubstituted hydrocarbons (which may form a substituted orunsubstituted hydrocarbon backbone), such as alkylene units;hydroxycarboxylic acid units (which may form a polyester backbone),e.g., glycolate, lactate, hydroxybutyrate, hydroxyvalerate,hydroxycaproate and hydroxybenzoate units; dicarboxylic acid units(which may form a polyester backbone in combination with a diol and/or apolyamide in combination with a diamine), e.g., adipate, succinate,terephthalate and naphthalene dicarboxylic acid units; diol units (whichmay form a polyether backbone, or form a polyester backbone incombination with a dicarboxylic acid), e.g., ethylene glycol,1,2-propanediol, 1,3-propanediol, 1,4-butanediol, and bisphenol A units;diamine units (which may form a polyamide backbone in combination with adicarboxylic acid), e.g., para-phenylene diamine and alkylene diaminessuch hexylene diamine; carbamate units (which may form a polyurethanebackbone); amino acid residues (which may form a polypeptide backbone);and saccharide residues (which may form a polysaccharide backbone).

In some embodiments of any of the embodiments described herein, Y is asubstituted or unsubstituted alkylene unit.

In some embodiments, Y is a substituted or unsubstituted ethylene unit,that is, an alkylene unit 2 atoms in length.

Polymeric backbones wherein Y is a substituted or unsubstituted ethyleneunit may optionally be a polymeric backbone such as formed bypolymerizing ethylene (CH₂═CH₂) and/or substituted derivatives thereof(also referred to herein as “vinyl monomers”). Such polymerization is avery well-studied procedure, and one of ordinary skill in the art willbe aware of numerous techniques for effecting such polymerization.

It is to be understood that any embodiments described herein relating toa polymeric backbone formed by a polymerization encompass any polymericbackbone having a structure which can be formed by such polymerization,regardless of whether the polymeric backbone was formed in practice bysuch polymerization (or any other type of polymerization).

As is well known in the art, the unsaturated bond of ethylene andsubstituted ethylene derivatives becomes saturated upon polymerization,such that the backbone units in a polymeric backbone are saturated,although they may be referred to as units of an unsaturated compound(e.g., a “vinyl monomer” or “olefin monomer”) to which they areanalogous.

Polymers which can be formed from unsaturated monomers such as vinylmonomers and olefin monomers are also referred to by the terms“polyvinyl” and “polyolefin”.

Herein, an “unsubstituted” alkylene unit (e.g., ethylene unit) refers toan alkylene unit which does not have any substituent other than thependant group discussed herein (represented as (-L-Z)). That is, analkylene unit attached to the aforementioned pendant group is consideredunsubstituted if there are no substituents at any other positions on thealkylene unit.

In some embodiments of any of the embodiments described herein, Y hasthe formula —CR₄R₅—CR₆D-.

When Y is a backbone unit which is not attached to L or Z (i.e., to apendant group described herein), D is R₇ (an end group, as definedherein); and when Y is a backbone unit which is attached to L or Z, D isa covalent bond or a linking group attaching Y to L or Z. The linkinggroup may optionally be —O—, —S—, arylene, sulfinyl, sulfonyl,phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea,thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, or amino.

R₄-R₇ are each independently hydrogen, alkenyl, alkynyl, cycloalkyl,aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro,azide, azo, phosphate phosphonyl, phosphinyl, oxo, carbonyl,thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl,N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, oramino.

Herein, the phrase “linking group” describes a group (e.g., asubstituent) that is attached to two or more moieties in the compound.

Herein, the phrase “end group” describes a group (e.g., a substituent)that is attached to a single moiety in the compound via one atomthereof.

When each of R₄-R₆ is hydrogen, and D is a covalent bond or linkinggroup, Y is an unsubstituted ethylene group attached (via D) to apendant group described herein.

When each of R₄-R₇ is hydrogen (and D is R₇), Y is an unsubstitutedethylene group which is not attached to a pendant group describedherein.

In some embodiments of any of the embodiments described herein, R₄ andR₅ are each hydrogen. Such embodiments include polymeric backbonesformed from many widely used vinyl monomers (including ethylene),including, for example, olefins (e.g., ethylene, propylene, 1-butylene,isobutylene, 4-methyl-1-pentene), vinyl chloride, styrene, vinylacetate, acrylonitrile, acrylate and derivatives thereof (e.g., acrylateesters, acrylamides), and methacrylate and derivatives thereof (e.g.,methacrylate esters, methacrylamides).

In some embodiments of any of the embodiments described herein, R₆ ishydrogen. In some such embodiments, R₄ and R₅ are each hydrogen.

In some embodiments of any of the embodiments described herein, R₆ ismethyl. In some such embodiments, R₄ and R₅ are each hydrogen. In somesuch embodiments, the backbone unit is a unit of methacrylate or aderivative thereof (e.g., methacrylate ester, methacrylamide).

In some embodiments of any of the embodiments described herein, thelinking group represented by the variable D is —O—, —C(═O)O—, —C(═O)NH—or phenylene. In exemplary embodiments, D is —C(═O)O—.

For example, the backbone unit may optionally be a vinyl alcoholderivative (e.g., an ester or ether of a vinyl alcohol unit) when D is—O—; an acrylate or methacrylate derivative (e.g., an ester of anacrylate or methacrylate unit) when D is —C(═O)O—; an acrylamide ormethacrylamide unit when D is —C(═O)NH—; and/or a styrene derivative(e.g., a substituted styrene unit) when D is phenylene.

In some embodiments of any of the embodiments described herein, L is asubstituted or unsubstituted hydrocarbon from 1 to 10 carbon atoms inlength. In some embodiments, the hydrocarbon is unsubstituted. In someembodiments, the hydrocarbon is a linear, unsubstituted hydrocarbon,that is, —(CH₂)_(i)— wherein i is an integer from 1 to 10.

In some embodiments of any of the embodiments described herein, L is asubstituted or unsubstituted ethylene group. In some embodiments, L isan unsubstituted ethylene group (—CH₂CH₂—).

In some embodiments of any of the embodiments described herein, B is anoxygen atom. In some such embodiments, L is a hydrocarbon according toany of the respective embodiments described herein (i.e., L is notabsent), and Z is a phosphate group attached to L.

In some embodiments of any of the embodiments described herein, B isabsent. In some such embodiments, L is a hydrocarbon according to any ofthe respective embodiments described herein (i.e., L is not absent), andZ is a phosphonate group attached to L. In some embodiments, L is alsoabsent, such that the phosphorus atom of formula II is attached directlyto Y.

In some embodiments of any of the embodiments described herein, A is asubstituted or unsubstituted hydrocarbon from 1 to 4 carbon atoms inlength.

In some embodiments of any of the embodiments described herein, A is anunsubstituted hydrocarbon. In some such embodiments, the unsubstitutedhydrocarbon is from 1 to 4 carbon atoms in length. In some embodiments,the hydrocarbon is a linear, unsubstituted hydrocarbon, that is,—(CH₂)_(i)— wherein j is an integer from 1 to 4.

In some embodiments of any of the embodiments described herein, A is asubstituted or unsubstituted ethylene group.

In some embodiments of any of the embodiments described herein, A is anunsubstituted ethylene group (—CH₂CH₂—). In such embodiments, the moietyhaving general formula II (represented by the variable Z) is similar oridentical to a phosphoethanolamine or phosphocholine moiety.Phosphoethanolamine and phosphocholine moieties are present in manynaturally occurring compounds (e.g., phosphatidylcholines,phosphatidylethanolamines).

In some embodiments of any of the embodiments described herein, A is anethylene group substituted by a C-carboxy group. In some embodiments,the C-carboxy is attached to the carbon atom adjacent to the nitrogenatom depicted in formula II (rather than the carbon atom attached to thedepicted oxygen atom). In such embodiments, the moiety having generalformula II (represented by the variable Z) is similar or identical to aphosphoserine moiety. Phosphoserine is present in many naturallyoccurring compounds (e.g., phosphatidylserines).

Without being bound by any particular theory, it is believed thatmoieties similar or identical to naturally occurring moieties such asphosphocholine, phosphoethanolamine and/or phosphoserine may beparticularly biocompatible.

In some embodiments of any of the embodiments described herein, R₁-R₃(the substituents of the nitrogen atom depicted in general formula II)are each independently hydrogen or C₁₋₄-alkyl. In some embodiments,R₁-R₃ are each independently hydrogen or methyl. In some embodiments,R₁-R₃ are each methyl. In some such embodiments, R₁-R₃ are eachhydrogen.

The variable n may be regarded as representing a number of backboneunits (represented by the variable Y) which are substituted by thependant group represented by (-L-Z), and the variable m may be regardedas representing a number of backbone units which are not substituted bysuch a pendant group. The sum n+m may be regarded as representing thetotal number of backbone units in the polymeric backbone. The ration/(n+m) may be regarded as representing the fraction of backbone unitswhich are substituted by the pendant group represented by (-L-Z).

The backbone unit Y substituted by the pendant group may be the same asor different than the backbone unit Y which is not substituted by thependant group (e.g., when m is at least 1).

The plurality (indicated by the variable n) of backbone units Ysubstituted by the pendant group may be the same as each other ordifferent from each other.

In addition, the plurality (indicated by the variable n) of pendantgroups (-L-Z) attached to a plurality of backbone units Y may be thesame as each other or different from each other (e.g., may differ in theidentity of any one or more of A, B, R₁, R₂, R₃ and L).

In any of the embodiments described herein wherein more than onebackbone unit Y is not substituted by the pendant group described herein(i.e., when m is more than 1), the plurality (indicated by the variablem) of backbone units Y which are substituted by the pendant group may bethe same as each other or different from each other.

The number of types of backbone units substituted by the pendant group,the number of types of backbone units not substituted by the pendantgroup (if any such units are present), and/or the number of types ofpendant group in the polymeric moiety may each independently be anynumber (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more).

In some embodiments of any of the embodiments described herein, thepolymeric moiety is a copolymer moiety, that is, the polymeric moietycomprises at least two different types of monomeric unit. The differenttypes of monomeric unit may differ in whether they comprise the pendantgroup (-L-Z) according to any of the respective embodiments describedherein (e.g., when m is at least 1), differ in the type of backbone unitY, and/or differ in the type of pendant group (-L-Z).

For example, in some embodiments of any of the embodiments describedherein the backbone unit Y in each of the Y(-L-Z) units may optionallybe the same or different, while the L and Z moieties are the same amongthe Y(-L-Z) units. In some such embodiments, backbone units notsubstituted by the pendant group (if any such units are present) mayoptionally be the same as backbone unit Y in each of the Y(-L-Z) units.Alternatively, backbone units not substituted by the pendant group (ifany such units are present) may optionally be different than backboneunit Y in each of the Y(-L-Z) units (while optionally being the sameamong all backbone units not substituted by the pendant group).

In some embodiments of any of the embodiments described herein the Lmoiety in each of the Y(-L-Z) units may optionally be the same ordifferent, while the backbone units Y and the Z moieties are the sameamong the Y(-L-Z) units. In some such embodiments, backbone units notsubstituted by the pendant group (if any such units are present) mayoptionally be the same as backbone unit Y in each of the Y(-L-Z) units.Alternatively, backbone units not substituted by the pendant group (ifany such units are present) may optionally be different than backboneunit Y in each of the Y(-L-Z) units (while optionally being the sameamong all backbone units not substituted by the pendant group).

In some embodiments of any of the embodiments described herein the Zmoiety in each of the Y(-L-Z) units may optionally be the same ordifferent, while the backbone units Y and the Z moieties are the sameamong the Y(-L-Z) units. In some such embodiments, backbone units notsubstituted by the pendant group (if any such units are present) mayoptionally be the same as backbone unit Y in each of the Y(-L-Z) units.Alternatively, backbone units not substituted by the pendant group (ifany such units are present) may optionally be different than backboneunit Y in each of the Y(-L-Z) units (while optionally being the sameamong all backbone units not substituted by the pendant group).

In any of the embodiments described herein wherein the polymeric moietyis a copolymer moiety, any two or more different types of monomeric unitmay be distributed randomly or non-randomly throughout the polymericmoiety. When different types of monomeric unit are distributednon-randomly, the copolymer may be one characterized by any non-randomdistribution, for example, an alternating copolymer, a periodiccopolymer, and/or a block copolymer.

In some embodiments of any of the embodiments described herein, at leasta portion of the monomeric units of the polymeric moiety comprise atargeting moiety (according to any of the embodiments described hereinrelating to a targeting moiety).

A targeting moiety may optionally be comprised by a backbone unit Yaccording to any of the respective embodiments described herein, linkingmoiety L according to any of the respective embodiments describedherein, and/or moiety Z according to any of the respective embodimentsdescribed herein, for example, wherein a substituent according to any ofthe respective embodiments described herein comprises (and optionallyconsists of) the targeting moiety. For example, in some embodimentswherein at least a portion of backbone units Y have the formula—CR₄R₅—CR₆D- (as described herein in any of the respective embodiments),any one or more of R₄-R₆ and D (optionally wherein D is R₇ as describedherein) comprises a targeting moiety according to any of the respectiveembodiments described herein (e.g., wherein any one or more of R₄-R₆ andD is a substituted group, comprising a substituent which is a targetingmoiety), and optionally any one or more R₄-R₆ and D is a targetingmoiety. However, many other structures of monomeric units comprising asubstituent which comprises (and optionally consist of) a targetingmoiety are also encompassed by embodiments of the invention.

When Y is a backbone unit which is not attached to L or Z (i.e., to apendant group as described herein), D is R₇ (an end group, as definedherein); and when Y is a backbone unit which is attached to L or Z, D isa covalent bond or a linking group attaching Y to L or Z. The linkinggroup may optionally be —O—, —S—, arylene, sulfinyl, sulfonyl,phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea,thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, or amino.

R₄-R₇ are each independently hydrogen, alkenyl, alkynyl, cycloalkyl,aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro,azide, azo, phosphate phosphonyl, phosphinyl, oxo, carbonyl,thiocarbonyl, urea, thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl,N-thiocarbamyl, C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, oramino.

In some embodiments, the polymeric moiety is a copolymer moiety whereinat least one type of monomeric unit comprises a targeting moiety(according to any of the respective embodiments described herein) and atleast one type of monomeric unit does not comprise such a targetingmoiety. The distribution of a monomeric unit comprising a targetingmoiety may be in accordance with any distribution described herein of amonomeric unit in a copolymer moiety (e.g., random, alternating,periodic copolymer, and/or block copolymer).

In some embodiments of any of the embodiments described herein wherein aportion of monomeric units comprise a targeting moiety, the monomericunits comprising a targeting moiety are, on average, closer to aterminus of the polymeric moiety distal to the lipid moiety, e.g., anaverage distance (as measured in atoms or backbone units along thebackbone of the polymeric moiety) of monomeric units comprising atargeting moiety from the lipid moiety is greater than an averagedistance of the other monomeric units from the lipid moiety.

In some embodiments, at least a portion (and optionally all) of themonomeric units comprising a targeting moiety form a block (of one ormore monomeric units) near (and optionally at) a terminus of thepolymeric moiety distal to the lipid moiety. In some such embodiments,the copolymer moiety contains a single monomeric unit which comprises atargeting moiety, and said monomeric unit is at a terminus of thepolymeric moiety distal to the lipid moiety.

Without being bound by any particular theory, it is assumed that atargeting moiety located distal to the lipid moiety may be moreeffective as a targeting moiety (e.g., more effective at binding to atarget), for example, due to the targeting moiety being less stericallyshielded (e.g., by a surface to which the lipid moiety is associated)and therefore more exposed to and thus better able to make contact withtargets in an aqueous environment.

In alternative embodiments, the polymeric moiety does not comprise atargeting moiety described herein according to any of the respectiveembodiments.

In some embodiments of any of the embodiments described herein, thepercentage of backbone units (represented by the variable Y) which aresubstituted by the pendant group represented by (-L-Z) (as representedby the formula 100%*n/(n+m)) is at least 20%. In some embodiments, thepercentage of backbone units substituted by the aforementioned pendantgroup is at least 30%. In some embodiments, the percentage of backboneunits substituted by the aforementioned pendant group is at least 40%.In some embodiments, the percentage of backbone units substituted by theaforementioned pendant group is at least 50%. In some embodiments, thepercentage of backbone units substituted by the aforementioned pendantgroup is at least 60%. In some embodiments, the percentage of backboneunits substituted by the aforementioned pendant group is at least 70%.In some embodiments, the percentage of backbone units substituted by theaforementioned pendant group is at least 80%. In some embodiments, thepercentage of backbone units substituted by the aforementioned pendantgroup is at least 90%. In some embodiments, the percentage of backboneunits substituted by the aforementioned pendant group is at least 95%.In some embodiments, the percentage of backbone units substituted by theaforementioned pendant group is at least 98%.

In some embodiments of any of the embodiments described herein, m is 0,such that each of the backbone units (represented by the variable Y) issubstituted by the pendant group represented by (-L-Z).

In some embodiments of any of the embodiments described herein, n is atleast 5. In some embodiments, n is at least 10. In some embodiments, nis at least 15.

In some embodiments of any of the embodiments described herein, n is ina range of from 2 to 1,000. In some embodiments of any of theembodiments described herein, n is in a range of from 2 to 500. In someembodiments of any of the embodiments described herein, n is in a rangeof from 2 to 200. In some embodiments of any of the embodimentsdescribed herein, n is in a range of from 2 to 100. In some embodimentsof any of the embodiments described herein, n is in a range of from 2 to50. In some such embodiments, m is 0.

In some embodiments of any of the embodiments described herein, n is ina range of from 3 to 1,000. In some embodiments of any of theembodiments described herein, n is in a range of from 3 to 500. In someembodiments of any of the embodiments described herein, n is in a rangeof from 3 to 200. In some embodiments of any of the embodimentsdescribed herein, n is in a range of from 3 to 100. In some embodimentsof any of the embodiments described herein, n is in a range of from 3 to50. In some embodiments of any of the embodiments described herein, n isin a range of from 5 to 50. In some embodiments of any of theembodiments described herein, n is in a range of from 10 to 50. In someembodiments of any of the embodiments described herein, n is in a rangeof from 10 to 25. In some such embodiments, m is 0.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 1,000. In some such embodiments, n is in a range offrom 2 to 1,000, such that the total number of backbone units is in arange of from 2 to 2,000. In some such embodiments, n is in a range offrom 3 to 1,000. In some embodiments, n is in a range of from 3 to 500.In some embodiments, n is in a range of from 3 to 200. In someembodiments, n is in a range of from 3 to 100. In some embodiments, n isin a range of from 3 to 50. In some embodiments, n is in a range of from5 to 50. In some embodiments, n is in a range of from 10 to 50.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 500. In some such embodiments, n is in a range offrom 2 to 1,000. In some such embodiments, n is in a range of from 3 to1,000. In some embodiments, n is in a range of from 3 to 500. In someembodiments, n is in a range of from 3 to 200. In some embodiments, n isin a range of from 3 to 100. In some embodiments, n is in a range offrom 3 to 50. In some embodiments, n is in a range of from 5 to 50. Insome embodiments, n is in a range of from 10 to 50.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 200. In some such embodiments, n is in a range offrom 2 to 1,000. In some such embodiments, n is in a range of from 3 to1,000. In some embodiments, n is in a range of from 3 to 500. In someembodiments, n is in a range of from 3 to 200. In some embodiments, n isin a range of from 3 to 100. In some embodiments, n is in a range offrom 3 to 50. In some embodiments, n is in a range of from 5 to 50. Insome embodiments, n is in a range of from 10 to 50.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 100. In some such embodiments, n is in a range offrom 2 to 1,000. In some such embodiments, n is in a range of from 3 to1,000. In some embodiments, n is in a range of from 3 to 500. In someembodiments, n is in a range of from 3 to 200. In some embodiments, n isin a range of from 3 to 100. In some embodiments, n is in a range offrom 3 to 50. In some embodiments, n is in a range of from 5 to 50. Insome embodiments, n is in a range of from 10 to 50.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 50. In some such embodiments, n is in a range offrom 2 to 1,000. In some such embodiments, n is in a range of from 3 to1,000. In some embodiments, n is in a range of from 3 to 500. In someembodiments, n is in a range of from 3 to 200. In some embodiments, n isin a range of from 3 to 100. In some embodiments, n is in a range offrom 3 to 50. In some embodiments, n is in a range of from 5 to 50. Insome embodiments, n is in a range of from 10 to 50.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 20. In some such embodiments, n is in a range offrom 2 to 1,000. In some such embodiments, n is in a range of from 3 to1,000. In some embodiments, n is in a range of from 3 to 500. In someembodiments, n is in a range of from 3 to 200. In some embodiments, n isin a range of from 3 to 100. In some embodiments, n is in a range offrom 3 to 50. In some embodiments, n is in a range of from 5 to 50. Insome embodiments, n is in a range of from 10 to 50.

In some embodiments of any of the embodiments described herein, m is ina range of from 0 to 10. In some such embodiments, n is in a range offrom 2 to 1,000. In some such embodiments, n is in a range of from 3 to1,000. In some embodiments, n is in a range of from 3 to 500. In someembodiments, n is in a range of from 3 to 200. In some embodiments, n isin a range of from 3 to 100. In some embodiments, n is in a range offrom 3 to 50. In some embodiments, n is in a range of from 5 to 50. Insome embodiments, n is in a range of from 10 to 50.

Lipid Moiety:

The lipid moiety (represented by the variable X in formula I herein)according to any of the embodiments in this section may be attached to apolymeric moiety according to any of the embodiments described in thesection herein relating to the polymeric moiety.

The lipid moiety may optionally be derived from any lipid known in theart (including, but not limited to, a naturally occurring lipid).Derivation of the lipid moiety from the lipid may optionally consist ofsubstituting a hydrogen atom at any position of the lipid with thepolymeric moiety represented in general formula I by [Y(-L-Z)]n[Y]m(i.e., the polymeric moiety represented by general formula Ia).

In some embodiments of any of the embodiments described herein, thelipid moiety (according to any of the respective embodiments describedherein) is attached to a Y(-L-Z) unit (according to any of theembodiments described herein relating to Y, L and/or Z), that is,backbone unit substituted by the pendant group described herein (e.g.,rather than a backbone unit not substituted by the pendant group).

Alternatively or additionally, in some embodiments of any of theembodiments described herein wherein m is at least 1, the lipid moiety(according to any of the respective embodiments described herein) mayoptionally be attached to a backbone unit (Y) which is not substitutedby a pendant group described herein (e.g., rather than attached to abackbone unit substituted by the pendant group). For example, thepolymeric moiety may optionally be a copolymer wherein the identity ofthe backbone unit attached to the lipid moiety varies randomly betweenmolecules. Thus, the depiction of X in Formula I as being attached to abackbone unit substituted by a pendant group (i.e., Y-(L-Z)) rather thanto an unsubstituted backbone unit Y is arbitrary, and is not intended tobe limiting.

In some embodiments of any of the embodiments described herein, thelipid moiety is a moiety of a lipid which is a fatty acid, amonoglyceride, a diglyceride, a triglyceride, a glycerophospholipid, asphingolipid, or a sterol. In some embodiments, the lipid is aglycerophospholipid.

In some embodiments of any of the embodiments described herein, thelipid moiety comprises at least one fatty acid moiety (e.g., an acylgroup derived from a fatty acid). The fatty acid moiety may be derivedfrom a saturated or unsaturated fatty acid. For example, the lipidmoiety may consist of a fatty acid moiety, or be a monoglyceride moietycomprising one fatty acid moiety, a diglyceride moiety comprising twofatty acid moieties, or a triglyceride moiety comprising three fattyacid moieties.

Examples of fatty acid moieties which may optionally be comprised by thelipid moiety include, without limitation, lauroyl, myristoyl, palmitoyl,stearoyl, palmitoleoyl, oleoyl, and linoleoyl.

Suitable examples of glycerophospholipids include, without limitation, aphosphatidyl ethanolamine, a phosphatidyl serine, a phosphatidylglycerol and a phosphatidyl inositol.

In some embodiments of any of the embodiments described herein, thelipid moiety represented by the variable X has the general formula III:

W₁ and W₂ are each independently hydrogen, alkyl, alkenyl, alkynyl oracyl, wherein at least one of W₁ and W₂ is not hydrogen;

J is —P(═O)(OH)—O—, or J is absent (such that K is attached directly tothe depicted oxygen atom of a glycerol moiety);

K is a substituted or unsubstituted hydrocarbon from 1 to 10 carbonatoms in length;

M is a linking group which is —O—, —S—, amino, sulfinyl, sulfonyl,phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea,thiourea, carbamyl, thiocarbamyl, amido, carboxy, or sulfonamide, or Mis absent (such that K is attached directly to Q); and

Q is a substituted or unsubstituted hydrocarbon from 1 to 10 carbonatoms in length, or Q is absent.

Q is attached to a backbone unit of the polymeric backbone according toany of the respective embodiments described herein, or alternatively,when Q is absent, M is attached to the aforementioned backbone unit.

When M is absent, Q is also absent, and K is attached to a backbone unitof the polymeric backbone according to any of the respective embodimentsdescribed herein.

In some embodiments of any of the embodiments described herein, one ofW₁ and W₂ is hydrogen and the other is not hydrogen.

In some embodiments of any of the embodiments described herein, neitherW₁ nor W₂ is hydrogen.

In some embodiments of any of the embodiments described herein, at leastone of W₁ and W₂ is an alkyl, alkenyl, alkynyl or acyl, which is from 10to 30 carbon atoms in length. In some embodiments, each of W₁ and W₂ isfrom 10 to 30 carbon atoms in length.

Examples of acyl groups which may optionally serve independently as W₁and/or W₂ include, without limitation, lauroyl, myristoyl, palmitoyl,stearoyl, palmitoleoyl, oleoyl, and linoleoyl.

In some embodiments of any of the embodiments described herein, J is—P(═O)(OH)—O— (e.g., the lipid moiety is a glycerophospholipid).

Herein, the length of the hydrocarbon represented by the variable Krefers to the number of atoms separating J and M (i.e., along theshortest path between J and M) as depicted in formula III.

When K is a substituted hydrocarbon, M may be attached to a carbon atomof the hydrocarbon per se, or be attached to a substituent of thehydrocarbon.

In some embodiments of any of the embodiments described herein, K is anacyl moiety (e.g., —C(═O)—C(CH₃)₂—). In some such embodiments, J isabsent, such that K is attached directly to the depicted oxygen atom ofa glycerol moiety. In some such embodiments, K comprises a carbonyllinking group (—C(═O)—), which attaches to the oxygen atom of a glycerolmoiety via an ester bond.

In some embodiments of any of the embodiments described herein, K is anethanolamine moiety (e.g., —CH₂—CH₂—NH—, or —CH₂—CH₂— attached to anitrogen atom), a serine moiety (e.g., —CH₂—CH(CO₂H)—NH—, or—CH₂—CH(CO₂H)— attached to a nitrogen atom), a glycerol moiety (e.g.,—CH(OH)—CH(OH)—CH—O—) and an inositol moiety (e.g.,-cyclohexyl(OH)₄—O—). In some embodiments, J is —P(═O)(OH)—O—.

In some embodiments of any of the embodiments described herein, M isamido, optionally —C(═O)NH—.

In some embodiments, the nitrogen atom of the amido is attached to K. Insome such embodiments, K is an ethanolamine or serine moiety describedherein.

In some embodiments of any of the embodiments described herein, Q is asubstituted or unsubstituted methylene group. In some such embodiments,M is amido. In some embodiments, the C(═O) of the amido is attached toQ.

In some embodiments of any of the embodiments described herein, Q is amethylene group substituted by one or two substituents. In someembodiments, the methylene group is substituted by one or two alkylgroups (e.g., C₁₋₄-alkyl).

In some embodiments of any of the embodiments described herein, Q is amethylene group substituted by two substituents. In some embodiments,the methylene group is substituted by two alkyl groups (e.g.,C₁₋₄-alkyl). In some embodiments, the alkyl groups are methyl, such thatQ is dimethylmethylene (—C(CH₃)₂—).

As exemplified in the Examples section herein, a substituted methylene(e.g., di-substituted methylene) represented by the variable Q isparticularly suitable for participating in polymerization reactions(e.g., as an initiator), because a free radical and/or ion on themethylene may be stabilized by the substituent(s) thereof.

As further exemplified herein, formation of an amido group (representedby the variable M) may serve as a convenient way to attach theabovementioned substituted methylene to a lipid (e.g., a naturallyoccurring lipid) such as a phosphatidylethanolamine orphosphatidylserine.

In some embodiments of any of the embodiments described herein, M and Qare each absent, and K is terminated by a substituted or unsubstitutedmethylene group, according to any of the respective embodimentsdescribed herein with respect to Q, for example, a methylene groupsubstituted by two substituents (e.g., dimethylmethylene (—C(CH₃)₂—)).In some embodiments, K further comprises a carbonyl group according toany of the respective embodiments described herein.

In some embodiments of any of the embodiments described herein, J, M andQ are each absent. In some such embodiments, K comprises a carbonyllinking group (—C(═O)—) attached directly to the depicted oxygen atom ofa glycerol moiety (via an ester bond), and further comprises asubstituted or unsubstituted methylene group (e.g., dimethylmethylene).In some embodiment, K consists of a carbonyl linking group attacheddirectly to the depicted oxygen atom of a glycerol moiety (via an esterbond), and a substituted or unsubstituted methylene group, for example,K is —C(═O)—C(CH₃)₂—.

Targeting Moiety:

As described hereinabove, in some embodiments of any of the embodimentsdescribed herein, at least a portion of the monomeric units comprise atargeting moiety (according to any of the embodiments described hereinrelating to a targeting moiety).

Herein, a “targeting moiety” refers to a moiety which is capable ofbringing a compound (e.g., a compound according to some embodiments ofthe invention) into proximity with a selected substance and/or material(which is referred to herein as a “target”). The target is optionally acell (e.g., a proliferating cell associated with the proliferativedisease or disorder), wherein the proximity is such that the targetingmoiety facilitates attachment and/or internalization of the compoundinto a target cell, and such that the compound may exert a therapeuticeffect.

In any of the embodiments described herein wherein m is at least 1, atleast a portion of the monomeric units comprising a targeting moiety(the number of which is represented by the variable m), according to anyof the respective embodiments described herein, are monomeric unitswhich do not comprise the pendant group represented by (-L-Z). In somesuch embodiments, each of the monomeric units comprising a targetingmoiety (according to any of the respective embodiments described herein)is a monomeric unit which comprises the pendant group represented by(-L-Z) (i.e., a backbone unit Y substituted by (-L-Z)), that is, none ofthe monomeric units comprising the pendant group represented by (-L-Z)comprise the aforementioned targeting moiety.

In any of the embodiments described herein wherein m is at least 1, eachof the monomeric units which do not comprise the pendant grouprepresented by (-L-Z) (the number of which is represented by thevariable m) comprises a targeting moiety (according to any of therespective embodiments described herein). In some such embodiments, eachof the monomeric units comprising a targeting moiety (according to anyof the respective embodiments described herein) is a monomeric unitwhich does not comprise the pendant group represented by (-L-Z), thatis, none of the monomeric units comprising the pendant group representedby (-L-Z) comprise the aforementioned targeting moiety, and each of themonomeric units which does not comprise the pendant group represented by(-L-Z) comprises the aforementioned targeting moiety.

In any of the embodiments described herein wherein m is at least 1, amonomeric unit comprising a targeting moiety may consist essentially ofa backbone unit Y (according to any of the respective embodimentsdescribed herein) substituted by one or more targeting moieties(according to any of the respective embodiments described herein).

The backbone unit Y of a monomeric unit comprising a targeting moietymay optionally be different (optionally considerably different) instructure than a backbone unit Y of other monomeric units in thepolymeric moiety (according to any of the respective embodimentsdescribed herein).

In any of the embodiments described herein wherein m is at least 1, thepolymeric moiety comprises a monomeric unit which comprises a targetingmoiety, and said monomeric unit is at a terminus of the polymeric moietydistal to the lipid moiety. In such embodiments, the compoundrepresented by general formula I has the formula Ib:

wherein:

T is a monomeric unit comprising a targeting moiety (according to any ofthe respective embodiments described herein);

X and T are attached to distal termini of the moiety represented by[Y(-L-Z)]n[Y]m−1; and

X, Y, L, Z, n and m are defined in accordance with any of theembodiments described herein relating to general formula I, with theproviso that m is at least 1.

It is to be understood that T in formula Ib is a type of monomeric unitrepresented by Y (i.e., without the pendant group represented by (-L-Z))in formulas I and Ia, and the number of monomeric units represented by Y(i.e., without the pendant group represented by (-L-Z)) other than T isrepresented by the value m−1, such that the total number of monomericunits without the pendant group represented by (-L-Z)), including T, isrepresented by the variable m, as in formulas I and Ia.

In some embodiments, m is 1, such that m−1 is zero, and the compoundrepresented by formula Ib consequently has the formula: X—[Y(-L-Z)]n-T,wherein L, T, X, Y, Z and n are defined in accordance with any of theembodiments described herein.

A monomeric unit comprising a targeting moiety according to any of therespective embodiments described herein may optionally be prepared bypreparing a monomer comprising a targeting moiety, and using saidmonomer to prepare a polymeric moiety described herein (e.g., bypolymerization of monomers according to any of the respectiveembodiments described herein) and/or by modifying a monomeric unit in apolymeric moiety subsequently to preparation of a polymeric moiety(e.g., by polymerization of monomers according to any of the respectiveembodiments described herein), using any suitable technique known in theart, including, but not limited to, techniques for conjugation.

In some embodiments of any of the embodiments described herein relatingto a targeting moiety, the targeting moiety does not comprise a moietyhaving general formula II (according to any of the respectiveembodiments described herein). For example, even if a moiety representedby general formula II is capable of forming a bond with a target asdescribed herein, the phrase “targeting moiety”, in some embodiments, isto be understood as relating to a moiety distinct from a moietyrepresented by variable Z (having general formula II).

In some embodiments of any one of the embodiments described herein, thependant group represented by (-L-Z) is selected so as not to form a bondwith the target and/or so as not to include a structure and/or propertyof a targeting moiety as described herein in any one of the respectiveembodiments. For example, in embodiments wherein a targeting moietycomprising a nucleophilic group (according to any of the respectiveembodiments described herein)—for example, an amine group—is capable offorming a bond (e.g., covalent bond) with a target, the variable Z(having general formula II) is optionally selected such that thedepicted amine/ammonium group is a tertiary amine/ammonium (i.e., nomore than one of R₁-R₃ is hydrogen) or quaternary ammonium (i.e., noneof R₁-R₃ is hydrogen), preferably a quaternary ammonium (e.g.,comprising a trimethylamino group, such as in phosphocholine). Tertiaryamine groups, and especially quaternary ammonium groups, may besignificantly less reactive nucleophilic groups than primary andsecondary amine groups.

In some embodiments of any of the embodiments described herein relatingto a targeting moiety, the targeting moiety comprises (and optionallyconsists of) at least one functional group capable of forming a covalentbond or non-covalent bond (preferably a selective non-covalent bond)with a substance and/or material (which is referred to herein as a“target”), e.g., at a surface of the target (e.g., a surface of a celland/or tissue).

Herein, the phrase “functional group” encompasses chemical groups andmoieties of any size and any functionality described herein (forexample, any functionality capable of forming a covalent bond ornon-covalent bond with a target).

A non-covalent bond according to any of the respective embodimentsdescribed herein may optionally be effected by non-covalent interactionssuch as, without limitation, electrostatic attraction, hydrophobicbonds, hydrogen bonds, and aromatic interactions.

In some embodiments, the targeting moiety comprises a functional groupcapable of forming a non-covalent bond which is selective for thetarget, e.g., an affinity (e.g., as determined based on a dissociationconstant) of the targeting moiety and/or functional group to the targetis greater than an affinity of the of the targeting moiety and/orfunctional group to most (or all) other compounds capable of forming anon-covalent bond with the targeting moiety.

In some embodiments of any one of the embodiments described herein, thefunctional group(s) are capable of forming a covalent bond with one ormore specific functional groups (e.g., hydroxy, amine, thiohydroxyand/or oxo groups) which are present on the target (e.g., a targetaccording to any of the respective embodiments described herein).

Examples of functional groups (in a targeting moiety) capable of forminga covalent bond with a target (according to any of the respectiveembodiments described herein) and the type of covalent bonds they arecapable of forming, include, without limitation:

nucleophilic groups such as thiohydroxy, amine (e.g., primary orsecondary amine) and hydroxy, which may form covalent bonds with, e.g.,a nucleophilic leaving group (e.g., any nucleophilic group describedherein), Michael acceptor (e.g., any Michael acceptor described herein),acyl halide, isocyanate and/or isothiocyanate (e.g., as describedherein) in a target;

nucleophilic leaving groups such as halo, azide (—N₃), sulfate,phosphate, sulfonyl (e.g. mesyl, tosyl), N-hydroxysuccinimide (NHS)(e.g. NHS esters), sulfo-N-hydroxysuccinimide, and anhydride, which mayform covalent bonds with, e.g., a nucleophilic group (e.g., as describedherein) in a target;

Michael acceptors such as enones (e.g., maleimide, acrylate,methacrylate, acrylamide, methacrylamide), nitro groups and vinylsulfone, which may form covalent bonds with, e.g., a nucleophilic group(e.g., as described herein) in a target, optionally thiohydroxy;

dihydroxyphenyl groups (according to any of the respective embodimentsdescribed herein), which may form covalent bonds with, e.g., anucleophilic group (e.g., as described herein) and/or a substituted orunsubstituted phenyl group (e.g., another dihydroxyphenyl group) in atarget, as described herein;

an acyl halide (—C(═O)-halogen), isocyanate (—NCO) and isothiocyanate(—N═C═S) group, which may form covalent bonds with, e.g., a nucleophilicgroup (e.g., as described herein) in a target;

a carboxylate (—C(═O)OH) group, which may form a covalent bond with,e.g., a hydroxyl group in a target to form an ester bond and/or an aminegroup (e.g., primary amine) in a target to form an amide bond(optionally by reaction with a coupling reagent such as a carbodiimide);and/or a carboxylate group is in a target and may form an amide or esterbond with an amine or hydroxyl group, respectively, in the targetingmoiety;

an oxo group (optionally in an aldehyde group (—C(═O)H)), which may forma covalent imine bond with an amine group (e.g., a primary amine) in atarget; and/or an oxo group (optionally in an aldehyde group) is in atarget and may form a covalent imine bond with an amine groups in thetargeting moiety; and/or

thiohydroxy groups, which may form covalent disulfide (—S—S—) bonds witha thiohydroxy group in a target.

Modification of a monomer (e.g., prior to polymerization) or a monomericunit of a polymeric moiety (e.g., subsequent to polymerization) tocomprise any of the functional groups described herein may optionally beperformed using any suitable technique for conjugation known in the art.The skilled person will be readily capable of selecting a suitabletechnique for any given molecule to be modified.

Herein, the term “dihydroxyphenyl” refers to an aryl group (as definedherein) which is a phenyl substituted by two hydroxyl groups at anypositions thereof. The phenyl may optionally be substituted byadditional substituents (which may optionally comprise additionalhydroxyl groups), to thereby form a substituted dihydroxyphenyl group;or alternatively, the phenyl comprises no substituents other than thetwo hydroxyl groups, such that the dihydroxyphenyl group is anunsubstituted dihydroxyphenyl group.

In some embodiments of any one of the embodiments described herein, thedihydroxyphenyl group is an ortho-dihydroxyphenyl (wherein the hydroxylgroups are attached to the phenyl at adjacent positions) or apara-dihydroxyphenyl (wherein the hydroxyl groups are attached toopposite sides of the phenyl ring), each being a substituted orunsubstituted dihydroxyphenyl. In some such embodiments, theortho-dihydroxyphenyl or para-dihydroxyphenyl is an unsubstituteddihydroxyphenyl.

A dihydroxyphenyl group according to any of the respective embodimentsdescribed herein may optionally bond covalently and/or non-covalently toa target according to any one or more attachment mechanism described fordihydroxyphenyl (catechol) groups in Lee et al. [PNAS 2006,103:12999-13003], Brodie et al. [Biomedical Materials 2011, 6:015014]and/or International Patent Application PCT/IL2015/050606, the contentsof each of which are incorporated in their entirety, and especiallycontents regarding bonds formed by dihydroxyphenyl (catechol) groups tosurfaces.

In some embodiments of any one of the embodiments described herein, thefunctional group capable of forming a bond to a target is a functionalgroup capable of forming a covalent bond with an amine group, optionallya primary amine group. In some such embodiments, the target comprises onor more amino acids or amino acid residues, for example, a peptide orpolypeptide of any length (e.g., at least two amino acid residues, forexample, proteins), and the amine groups may optionally be lysine sidechain amine groups and/or N-terminal amine groups. In some embodiments,the target comprises an extracellular matrix protein, for example,collagen. In some embodiments, the target comprises cartilage (e.g.,articular cartilage).

In some embodiments of any one of the embodiments described herein, thetargeting moiety comprises (and optionally consists of) at least onefunctional group capable of forming a non-covalent bond with the target(e.g., as described herein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein, afunctional group capable of forming a non-covalent bond with the targetcomprises (and optionally consists of) a polysaccharide and/orpolypeptide (e.g., a protein and/or fragment thereof), wherein thetarget optionally comprises a ligand of the polysaccharide and/orpolypeptide; and/or the target comprises a polysaccharide and/orpolypeptide (e.g., a protein and/or fragment thereof) and the functionalgroup capable of forming a non-covalent bond with the target is a ligandof the polysaccharide and/or polypeptide.

Examples of suitable polysaccharides and/or polypeptides, and ligandsthereof, include, without limitation:

avidin or streptavidin as a polypeptide described herein, and biotin asa ligand thereof;

a polysaccharide-binding polypeptide as a polypeptide described therein,and a complementary polysaccharide as a ligand thereof (or acomplementary polysaccharide-binding polypeptide as a ligand of apolysaccharide described herein);

a collagen-binding polypeptide as a polypeptide described therein, and acomplementary collagen as a ligand thereof (or a collagen as apolypeptide described herein and a complementary collagen-bindingpolypeptide as a ligand thereof);

a cell receptor expressed by a cell, and a ligand selectively bound bythe receptor;

an antibody towards any antigen (e.g., wherein the target describedherein optionally comprises the antigen) or a fragment of such anantibody as a polypeptide described herein, and the respective antigenas a ligand thereof; and

an antibody mimetic towards any antigen (e.g., wherein the targetdescribed herein optionally comprises the antigen).

Examples of cell receptors expressed by a cell include, withoutlimitation, receptors characteristic of a particular type of cell and/ortissue, and receptors overexpressed by a cancer cell. The cell receptoror the cell is optionally a target described herein, and the targetingmoiety optionally comprises any ligand of the receptor. Examples of suchligands include, without limitation, transferrin, a ligand oftransferrin receptor which may optionally target transferrin receptoroverexpressed by some cancer cells; keratinocyte growth factor (KGF orFGF7) which is specific for cells of epithelial origin, and mayoptionally target KGF receptor such as that overexpressed by anendometrial carcinoma or pancreatic carcinoma [Visco et al., Int J Oncol1999, 15:431-435; Siegfried et al., Cancer 1997, 79:1166-1171]; andepidermal growth factor (EGF) which may optionally target an EGFreceptor, optionally an erbB, such as that overexpressed by gliomas andendometrial carcinomas [Normanno et al., Curr Drug Targets 2005,6:243-257]).

As used herein, the term “antibody” encompasses any type ofimmunoglobin. As used herein, the phrase “antibody mimetic” encompassesany type of molecule, optionally a polypeptide, referred as such in theart capable of selectively binding an antigen (e.g., non-covalently).Non-limiting examples of antibody mimetics include affibodies, affilins,affimers, affitins, alphabodies, anticalins, avimers, DARPins, Fynomers,Kunitz domain peptides, and monobodies, e.g., as described in Nygren[FEBS J 2008, 275:2668-2676], Ebersbach et al. [J Mol Biol 2007,372:172-185], Johnson et al. [Anal Chem 2012, 84:6553-6560], Krehenbrinket al. [J Mol Biol 2008, 383:1058-1068], Desmet et al. [Nature Comm2014, 5:5237], Skerra [FEBS J 2008, 275:2677-2683], Silverman et al.[Nature Biotechnol 2005, 23:1556-1561], Stumpp et al. [Drug Discov Today2008, 13:695-701], Grabulovski et al. [J Biol Chem 2007, 282:3196-3204],Nixon & Wood [Curr Opin Drug Discov Devel 2006, 9:261-268], Koide &Koide [Methods Mol Biol 2007, 325:95-109], and Gebauer & Skerra [CurrOpin Chem Biol 2009, 13:245-255], the contents of each of which areincorporated in their entirety, and especially contents regardingparticular types of antibody mimetics.

As used herein, the phrase “polysaccharide-binding polypeptide”encompasses any polypeptide or oligopeptide (peptide chains of at least2, and preferably at least 4 amino acid residues in length) capable ofselectively binding (e.g., non-covalently) to a polysaccharide. A widevariety of polysaccharide-binding polypeptides and their bindingspecificities will be known to the skilled person, and include shortpeptide sequences (e.g., from 4 to 50, optionally 4 to 20 amino acidresidues in length), and longer polypeptides such as proteins orfragments (e.g., carbohydrate-binding modules and/or domains) thereof.In addition, the phrase “polysaccharide-binding polypeptide” encompassesantibodies capable of specifically binding to a polysaccharide. Suchantibodies will be available to the skilled person and/or the skilledperson will know how to prepare such antibodies, using immunologicaltechniques known in the art.

Examples of polysaccharide-binding polypeptides which may be used insome of any one of the embodiments of the invention include, withoutlimitation, carbohydrate-binding modules (CBMs); and hyaluronicacid-binding peptides, polypeptides and/or modules (e.g., having asequence as described in any of International Patent Applicationpublication WO 2013/110056; International Patent Application publicationWO 2014/071132; Barta et al. [Biochem J 1993, 292:947-949], Kohda et al.[Cell 1996, 86:767-775], Brisset & Perkins [FEBS Lett 1996,388:211-216], Peach et al. [J Cell Biol 1993, 122:257-264], Singh et al.[Nature Materials 2014, 13:988-995], and Zaleski et al. [AntimicrobAgents Chemother 2006, 50:3856-3860], the contents of each of which areincorporated in their entirety, and especially contents regardingparticular polysaccharide-binding polypeptides), for example,GAHWQFNALTVR (a hyaluronic acid-binding peptide sequence).

Examples of CBMs which may be used in some of any one of the embodimentsof the invention, include, without limitation, CBMs belonging to thefamilies CBM3, CBM4, CBM9, CBM10, CBM17 and/or CBM28 (which mayoptionally be used to bind cellulose, e.g., in a cellulose-containingtarget); CBM5, CBM12, CBM14, CBM18, CBM19 and/or CBM33 (which mayoptionally be used to bind chitin and/or other polysaccharidescomprising N-acetylglucosamine, e.g., in a chitin-containing target);CBM15 (which may optionally be used to bind hemicellulose, e.g., in ahemicellulose-containing target); and/or CBM20, CBM21 and/or CBM48(which may optionally be used to bind starch and/or glycogen, e.g., in astarch-containing and/or glycogen-containing target).

As used herein, the phrase “collagen-binding polypeptide” encompassesany polypeptide or oligopeptide (peptide chains of at least 2, andpreferably at least 4 amino acid residues in length) capable ofselectively binding (e.g., non-covalently) to a collagen (e.g., one typeof collagen, some types of collagen, all types of collagen), includingglycosylated polypeptides and oligopeptides such as peptidoglycans andproteoglycans. A wide variety of collagen-binding polypeptides and theirbinding specificities will be known to the skilled person, and includeshort peptide sequences (e.g., from 4 to 50, optionally 4 to 20 aminoacid residues in length), and longer polypeptides such as proteins orfragments (e.g., collagen-binding domains) thereof. In addition, thephrase “collagen-binding polypeptide” encompasses antibodies capable ofspecifically binding to a collagen. Such antibodies will be available tothe skilled person and/or the skilled person will know how to preparesuch antibodies, using immunological techniques known in the art.

Examples of collagen-binding polypeptides which may be used inembodiments of the invention include, without limitation,collagen-binding proteins (e.g., decorin), fragments thereof and/orother polypeptides as described in U.S. Pat. No. 8,440,618, Abd-Elgaliel& Tung [Biopolymers 2013, 100:167-173], Paderi et al. [Tissue Eng Part A2009, 15:2991-2999], Rothenfluh et al. [Nat Mater 2008, 7:248-254] andHelms et al. [J Am Chem Soc 2009, 131:11683-11685] (the contents of eachof which are incorporated in their entirety, and especially contentsregarding particular collagen-binding polypeptides), for example, thesequence WYRGRL.

It is expected that during the life of a patent maturing from thisapplication many relevant functional groups and moieties for bindingwill be developed and/or uncovered and the scope of the terms “targetingmoiety”, “functional group”, “cell receptor”, “antibody”, “antibodymimetic”, “collagen-binding polypeptide” and “polysaccharide-bindingpolypeptide” and the like is intended to include all such newtechnologies a priori.

In some embodiments of any of the embodiments described herein, afunctional group in a targeting moiety (according to any of therespective embodiments described herein) is attached to a linking group(as defined herein). The linking group may optionally be any linkinggroup or linking moiety described herein, including, without limitation,a substituted or unsubstituted hydrocarbon. In some embodiments, thetargeting moiety (optionally a substituent of a backbone unit Y)consists essentially of a functional group attached to the rest of thepolymeric moiety via the linking group.

A functional group may optionally be attached to the linking moiety by acovalent bond obtainable by a reaction between two functional groups,for example, any covalent bond and/or functional groups described hereinin the context of forming a covalent bond between a functional group anda target.

In some embodiments of any of the embodiments described herein relatingto a functional group comprising a peptide or polypeptide, an amino acidresidue of the peptide or polypeptide is optionally attached to alinking group of the targeting moiety, for example, via an amide bondformed from an amine or carboxylate group in the peptide or polypeptide(e.g., in an N-terminus, a lysine side chain, a C-terminus, a glutamateside chain and/or an aspartate side chain), an ester bond formed from ahydroxyl or carboxylate group in the peptide or polypeptide (e.g., in aserine side chain, a threonine side chain, a C-terminus, a glutamateside chain and/or an aspartate side chain), and/or a disulfide bondformed from a thiohydroxy group in the peptide or polypeptide (e.g., ina cysteine side chain). In some embodiments, an amino acid residueattached to the linking group is an N-terminal and/or C-terminalresidue, for example, any amino acid residue attached via an N-terminalamino group or C-terminal carboxylate group, and/or a terminal lysine,glutamate, aspartate, serine, threonine and/or cysteine residue attachedvia a side chain thereof.

In some embodiments, an amino acid residue and/or peptide (e.g., from 2to 20 amino acid residues in length) is added to the N-terminus and/orC-terminus of a peptide or polypeptide sequence of a functional group(according to any of the respective embodiments described herein), andlinks the aforementioned sequence to a linking group. Examples of suchterminal amino acid residues and/or peptides include, withoutlimitation, glycine residues and peptides with a terminal glycineresidue, which may be used to attach a linking group to an N-terminus orC-terminus (according to any of the respective embodiments describedherein); serine and threonine residues and peptides with a terminalserine or threonine residue, which may be used to attach a linking groupto hydroxyl group in a serine or threonine side chain, optionally via anester bond (according to any of the respective embodiments describedherein); and cysteine residues and peptides with a terminal cysteineresidue, which may be used to attach a linking group to a peptide via adisulfide bond (according to any of the respective embodiments describedherein).

In some embodiments, attachment of a peptide or polypeptide to a linkinggroup via a terminal amino acid residue minimizes interference (e.g.,steric interference) with the functionality of the peptide orpolypeptide following attachment to the linking group.

In some embodiments, attachment of a peptide or polypeptide to a linkinggroup via a terminal glycine facilitates attachment by minimizinginterference (e.g., steric interference) of an amino acid side chain(which glycine lacks) with attachment to the linking group.

Lipid Layers and Liposomes:

According to an aspect of some embodiments of the invention, there isprovided a lipid bilayer (referred to herein interchangeably as simply a“bilayer”) comprising a polymeric compound according to any of therespective embodiments described herein. In some such embodiments, thebilayer further comprises at least one bilayer-forming lipid in additionto the polymeric compound.

Herein, the term “bilayer-forming lipid” encompasses any compound inwhich a bilayer may form from a pure aqueous solution of the compound,the bilayer comprising two parallel layers of molecules of the compound(referred to as a “lipid”).

Typically, the bilayer comprises relatively polar moieties of the lipidat the two surfaces of the bilayer, which may optionally comprise aninterface with the aqueous solution and/or an interface with a solidsurface; and relatively hydrophobic moieties of the lipid at theinterior of the bilayer, at an interface between the two layers of lipidmolecules which form the bilayer.

Examples of bilayer-forming lipids include glycerophospholipids (e.g., aglycerophospholipid according to any of the respective embodimentsdescribed herein). It is to be appreciated that the polymeric compounddescribed herein may optionally be a bilayer-forming lipid which canform a bilayer per se or in combination with one or more additionalbilayer-forming lipids.

In some embodiments of any one of the embodiments described herein, thebilayer-forming lipid comprises at least one charged group (e.g., one ormore negatively charged groups and/or one or more positively chargedgroups).

In some embodiments, the bilayer-forming lipid is zwitterionic;comprising both (e.g., an equal number of) negatively charged andpositively charged groups (e.g., one of each).

In some embodiments of any of the embodiments described herein relatingto a bilayer, a molar ratio of the bilayer-forming lipid (comprised inaddition to the polymeric compound) and the polymeric compound in thebilayer is in a range of from 5:1 to 5,000:1 (bilayer-forminglipid:polymeric compound), optionally in a range of from 10:1 to2,500:1, optionally in a range of from 25:1 to 1,000:1, and optionallyin a range of from 50:1 to 500:1.

In some embodiments of any of the embodiments described herein relatingto a bilayer, a polymeric moiety in the bilayer comprises a lipid moietyrepresented by the variable X in formula I (according to any of therespective embodiments described herein) which comprises a residue of abilayer-forming lipid (e.g., a glycerophospholipid) which is comprisedby the bilayer in addition to the polymeric moiety or which is closelyrelated to a bilayer-forming lipid comprises by the bilayer, forexample, wherein the lipid moiety comprises a glycerophospholipidresidue and the bilayer comprises another glycerophospholipid as abilayer-forming lipid (e.g., optionally wherein fatty acid residues inthe glycerophospholipid residue have about the same length as fatty acidresidues in the bilayer-forming lipid, and optionally wherein the fattyacid residues in the glycerophospholipid residue are substantially thesame as the fatty acid residues in the bilayer-forming lipid).

Without being bound by any particular theory, it is believed thatsimilarity between a lipid moiety of a polymeric moiety and abilayer-forming lipid facilitates anchorage of the lipid moiety of thepolymeric moiety in a bilayer comprising the bilayer-forming lipid.

The bilayer according to embodiments described herein may optionally beclosed upon itself (e.g., such that the bilayer has no edges), therebyforming an inner volume separated by the bilayer from the surroundingenvironment, which is referred to herein and in the art as a “liposome”.Alternatively or additionally, the bilayer may be open-faced and/or withedges.

According to an aspect of some embodiments of the invention, there isprovided a liposome comprising at least one lipid bilayer according toany of the respective embodiments described herein.

A liposome may optionally comprise a single bilayer or a plurality ofbilayers (each bilayer optionally independently forming a closedvesicle) comprising, for example, concentric bilayer vesicles and/or aplurality of separate bilayer vesicles encompassed by the same bilayervesicle.

A liposome according to any of the respective embodiments describedherein may be approximately spherical in shape or may have anyalternative shape, such as an elongated tube and/or a flattened (e.g.,sheet-like) shape.

In some embodiments of any of the embodiments described herein relatingto a liposome, the liposome further comprises at least one functionalmoiety or agent bound to a surface of the liposome and/or within a lipidbilayer and/or core of the liposome (e.g., within the liposome bilayerand/or enveloped by the liposome bilayer).

Examples of functional moieties and agents suitable for inclusion inembodiments described herein include, without limitation, atherapeutically active agent or moiety of a therapeutically active agent(e.g., wherein the active agent is releasable upon cleavage of themoiety), a labeling moiety or agent, and/or a targeting moiety ortargeting agent (e.g., a targeting moiety or agent on a surface of theliposome).

Examples of therapeutically active agents suitable for inclusion in aliposome (e.g., as a molecule or moiety of the agent) include, withoutlimitation, amphotericin B, cisplatin, cytarabine, daunorubicin,doxorubicin, estradiol, influenza virosome, morphine, surfactant proteinB, surfactant protein C, verteporfin and vincristine.

Examples of a labeling moiety or agent include moieties and compoundswhich are chromophoric (e.g., absorb visible light), fluorescent,phosphorescent, and/or radioactive. Many such compounds and moieties(and techniques for preparing such moieties) will be known to a skilledperson.

A targeting moiety in a liposome according to any of the respectiveembodiments described herein may optionally be a targeting moietyaccording to any of the respective embodiments described herein. Atargeting moiety in a liposome may be comprised by a polymeric compoundaccording to some embodiments of the invention (according to any of therespective embodiments described herein), the liposome comprising thepolymeric compound. Alternatively or additionally, a targeting moiety ina liposome may optionally be comprised by another compound in theliposome, optionally a bilayer-forming lipid (according to any of therespective embodiments described herein) conjugated to a targetingmoiety according to any of the respective embodiments described herein.

Herein, a “targeting agent” refers to a compound (“agent”) comprising(and optionally consisting essentially of) a targeting moiety accordingto any of the respective embodiments described herein (e.g., in thecontext of a targeting moiety comprised by a polymeric compounddescribed herein). Typically, the phrase “targeting agent” is used torefer to a compound other than a polymeric compound comprising atargeting moiety, as described herein.

In some embodiments, a functional moiety (e.g., targeting moiety orlabeling moiety) is covalently attached to a liposome. Such attachmentmay be obtained in some embodiments by using techniques known in the art(e.g., amide bond formation).

According to another aspect of embodiments of the invention, there isprovided a composition-of-matter comprising a substrate coated, on atleast a portion of a surface thereof, by at lipid bilayer according toany of the respective embodiments described herein.

According to another aspect of embodiments of the invention, there isprovided an article of manufacture comprising a composition-of-matteraccording to any one of the embodiments described herein.

Herein, the term “composition-of-matter” refers to any compositioncomprising a plurality of substances (e.g., substrate, water-solublepolymer(s), and amphiphilic lipid) in a form which does not exist innature, and which does not include a portion of a human being. The formwhich does not exist in nature may optionally comprise naturalsubstances in a combination which does not exist in nature, and/or mayoptionally comprise one or more substances which do not occur in nature.It is to be understood that this definition is not necessarily identicalwith a standard legal definition of the term.

Herein, the term “article of manufacture” refers to any article producedfrom materials in a manner which results in new forms, qualities,properties or combinations of the materials. It is to be understood thatthis definition is not necessarily identical with a standard legaldefinition of the term. The article of manufacture described herein mayoptionally consist essentially of the composition-of-matter, oralternatively, may comprise additional materials and/or parts.

At least a portion of the molecules of the amphiphilic lipid areoriented such that polar groups thereof (e.g., charged groups) faceoutwards at a surface of the composition-of-matter.

As used herein, the phrase “face outwards at a surface” refers to agroup in a molecule (e.g., a lipid) which is closer to the surface ofthe composition-of-matter than the center of gravity of the molecule isto the surface of the composition-of-matter, and farther from thesubstrate than the center of gravity of the molecule is from thesubstrate.

As discussed herein, and without being bound by any particular theory,it is believed that outwards facing polar groups (e.g., charged groups)according to some embodiments of the invention are effect highlyeffective lubrication and/or inhibition of adhesion, biofouling and/orbiofilm formation (e.g., as described herein) due, at least in part, toproperties of hydrated polar groups (e.g., hydration lubrication),especially hydrated charged groups.

In any of the embodiments described herein, the substrate may compriseany type of material or combination of different types of material,including inorganic material and/or organic material, in crystalline,amorphous and/or gel (e.g., hydrogel) forms, for example, metal,mineral, ceramic, glass, polymer (e.g., synthetic polymer, biopolymer),plant and/or animal biomass, and combinations thereof.

In some embodiments, the substrate comprises a physiological surface(e.g., a physiological tissue) and/or a surface in contact with and/orintended to come into contact with a physiological surface (e.g., asdescribed herein in any one of the respective embodiments).

In some embodiments of any of the embodiments described herein, thearticle is a medical device (the substrate being a medical device orportion of a medical device, as described herein). In some embodiments,the medical device is a device designed to come into contact with a partof the body susceptible to infection, such as an internal portion of thebody, a mucous membrane and/or a surface of the eye. Examples of suchmedical devices include, without limitation, surgical tools and implants(which are for coming into contact with an internal portion of the body)and contact lenses (which are for contacting a surface of the eye).

As used herein throughout, the phrase “medical device” includes anymaterial or device that is used on, in, or through a subject's body, forexample, in the course of medical treatment (e.g., for a disease orinjury). The subject may be human or a non-human animal, such that thephrase “medical device” encompasses veterinary devices. Medical devicesinclude, but are not limited to, such items as medical implants(including permanent implants and transient implants), wound caredevices, medical devices for drug delivery, contact lenses and bodycavity and personal protection devices. The medical implants include,but are not limited to, catheters (e.g., urinary catheters,intravascular catheters), injection ports, intubation equipment,dialysis shunts, wound drain tubes, skin sutures, vascular grafts,implantable meshes, intraocular devices, heart valves, and the like.Wound care devices include, but are not limited to, general wounddressings, biologic graft materials, tape closures and dressings, andsurgical incise drapes. Medical devices for drug delivery include, butare not limited to, needles, drug delivery skin patches, drug deliverymucosal patches and medical sponges. Body cavity and personal protectiondevices, include, but are not limited to, tampons, sponges, surgical andexamination gloves, and toothbrushes. Birth control devices include, butare not limited to, intrauterine devices (IUDs), diaphragms and condoms.

In the context of medical devices, it is to be understood that themedical device is coated by a bilayer described herein, and a bilayer orbilayer-comprising liposome per se is not considered herein to be amedical device.

Exemplary articles include the following:

Medical devices such as, but not limited to, pacemakers, heart valves,replacement joints, catheters, catheter access ports, dialysis tubing,gastric bands, shunts, screw plates, artificial spinal discreplacements, internal implantable defibrillators, cardiacresynchronization therapy devices, implantable cardiac monitors, mitralvalve ring repair devices, left ventricular assist devices (LVADs),artificial hearts, implantable infusion pumps, implantable insulinpumps, stents, implantable neurostimulators, maxillofacial implants,dental implants, and the like;

Packages or containers, for example, food packages and containers,beverage packages and containers, medical device packages, agriculturalpackages and containers (of agrochemicals), blood sample or otherbiological sample packages and containers, and any other packages orcontainers of various articles;

Food packages such as packages of dairy products and/or containers forstorage or transportation of dairy products;

Milk storage and processing devices such as, but not limited to,containers, storage tanks, raw milk holding equipment, dairy processingoperations conveyer belts, tube walls, gaskets, rubber seals, stainlesssteel coupons, piping systems, filling machine, silo tanks, heatexchangers, post-pasteurization equipment, pumps, valves, separators,and spray devices;

Energy harvesting device, for example, a microelectronic device, amicroelectromechanical device, a photovoltaic device and the like;

Microfluidic devices, for example, micro-pumps or micro valves and thelike;

Sealing parts, for example, 0 rings, and the like;

Articles having a corrodible surface;

Agricultural devices, as, for example, described herein;

Textiles, for example, tough cottons;

Fuel transportation devices;

Construction elements, such as, but not limited to, paints, walls,windows, door handles, and the like;

Elements of water treatment systems (such as for containing and/ortransporting and/or treating aqueous media or water), devices,containers, filters, tubes, solutions and gases and the like; and

Elements of organic waste treatment systems (such as for containingand/or disposing and/or transporting and/or treating organic waste),devices, containers, filters, tubes, solutions and gases and the like.

Lubrication:

Liposomes and bilayers described herein may optionally be useful forlubricating a surface, for example, a surface coated by a bilayerdescribed herein, and/or contacted with liposomes described herein.

According to an aspect of some embodiments of the invention, there isprovided a lubricant composition comprising liposomes according to anyof the respective embodiments described herein.

Herein, a “lubricant composition” refers to a composition intended forreducing a friction coefficient of a surface (e.g., according to amethod described herein).

In some embodiments, the lubricant composition comprises a carrier. Thecarrier may optionally be a liquid carrier. In some embodiments, thecarrier comprises an aqueous liquid.

In some embodiments, the lubricant composition (or any other compositiondescried herein comprising liposomes) further comprises a water-solublepolymer, optionally as part of the carrier.

As used herein, the phrase “water-soluble polymer” encompasses polymershaving a solubility of at least 1 gram per liter in an aqueous (e.g.,water) environment at pH 7 (at 25° C.).

In some embodiments of any of the embodiments described herein, thewater-soluble polymer has a solubility of at least 2 grams per liter(under the abovementioned conditions). In some embodiments, thesolubility is at least 5 grams per liter. In some embodiments, thesolubility is at least 10 grams per liter. In some embodiments, thesolubility is at least 20 grams per liter. In some embodiments, thesolubility is at least 50 grams per liter. In some embodiments, thesolubility is at least 100 grams per liter.

The water-soluble polymer(s) according to any of the embodimentsdescribed herein may comprise at least one ionic polymer and/or at leastone non-ionic polymer which are water-soluble as defined herein.

As used herein, the phrase “non-ionic polymer” refers to a polymer whichdoes not have a charged group.

Examples of suitable non-ionic water-soluble polymers include, withoutlimitation, polyvinylpyrrolidone (also referred to hereininterchangeably as povidone and/or PVP) and polyethylene oxide (alsoreferred to herein interchangeably as PEO, PEG and/or polyethyleneglycol).

As used herein, the phrase “ionic polymer” refers to polymers having atleast one charged group, and encompasses polymers having a net negativecharge (also referred to herein as “anionic polymers”), polymers havinga net positive charge (also referred to herein as “cationic polymers”),and polymers having no net charge (also referred to herein as“zwitterionic polymers”), in an aqueous (e.g., water) environment at pH7.

Herein throughout, the phrase “charged group” refers to any functionalgroup (e.g., a functional group described herein) which is ionic (asdefined herein), including, for example, amine, carboxylic acid,sulfate, sulfonate, phosphate and phosphonate. Thus, each electriccharge in a moiety or molecule is associated with one charged group,although a single charged group (e.g., non-substituted phosphate) may beassociated with more than one electric charge of the same sign (e.g., adianion, a dication).

Herein throughout, the term “ionic” refers to the presence of anelectric charge on at least one atom in a moiety and/or molecule (in atleast 50% of moieties and/or molecules in a population) in an aqueousmedium (e.g., water) at pH 7. The electric charge may be negative(anionic) or positive (cationic). If more than one electric charge ispresent, the electric charges may be negative (anionic) and/or positive(cationic), for example, both a negative and a positive charge may bepresent (zwitterionic).

Examples of ionic polymers include, without limitation, ionicpolysaccharides, such as hyaluronic acid, chondroitin sulfate, alginicacid, xanthan gum, chitosan and N-alkyl chitosan derivatives.

According to another aspect of embodiments described herein, there isprovided a method of reducing a friction coefficient of a surface, themethod comprising contacting the surface with liposomes according to anyof the respective embodiments described herein. In some embodiments, themethod is effected by contacting the surface with a compositioncomprising the liposomes and a carrier (optionally a lubricantcomposition according to any of the respective embodiments describedherein).

In some of any one of the embodiments described herein which relate to alubrication, according to any one of the aspects described herein, thelubrication is optionally effected according to any of the embodimentsdescribed in International Patent Application PCT/IL2015/050605 and/orPCT/IL2015/050606, which are incorporated herein by reference(especially in respect to methods and compositions for lubricating asurface), with the proviso that at least a portion of the liposomes usedare in accordance with any of the embodiments described herein.

In some embodiments, the method further comprises contacting the surfacewith a water-soluble polymer (e.g., according to any of the respectiveembodiments described herein), optionally prior to and/or concomitantlywith contacting the surface with liposomes. In some embodiments, themethod is effected by contacting the surface with a compositioncomprising the liposomes and the water-soluble polymer (optionally alubricant composition comprising the water-soluble polymer according toany of the respective embodiments described herein), optionally incombination with an aqueous liquid.

In some of any one of the embodiments described herein which relate to amethod and/or lubrication composition for reducing a frictioncoefficient of a surface, the surface is a hydrogel surface. In someembodiments, the hydrogel consists essentially of a polymer and anaqueous liquid (optionally water).

In some of any one of the embodiments described herein which relate to amethod and/or lubrication composition for reducing a frictioncoefficient of a surface, the surface is a contact lens surface.

In some of any one of the embodiments described herein which relate to acontact lens, according to any one of the aspects described herein, thecontact lens comprises a hydrogel surface. In some embodiments, thecontact lens comprises a hydrogel surface and a rigid center. In someembodiments, the contact lens consists essentially of a hydrogel.

The hydrogel may comprise any material known in the art for use incontact lens hydrogels. Examples of such hydrogel materials include,without limitation, alphafilcon A, asmofilcon A, balafilcon A, bufilconA, comfilcon A, crofilcon, deltafilcon A, dimefilcon, droxifilcon A,enfilcon A, etafilcon A, galyfilcon A, hefilcon A, hefilcon B,hilafilcon A, hilafilcon B, hioxifilcon A, hioxifilcon D, isofilcon,lidofilcon A, lidofilcon B, lotrafilcon B, mafilcon, methafilcon A,methafilcon B, narafilcon A, narafilcon B, ocufilcon A, ocufilcon B,ofilcon A, omafilcon A, perfilcon, phemfilcon A, polymacon, scafilcon A,senofilcon A, surfilcon, tefilcon, tetrafilcon A, tetrafilcon B,vifilcon A, and xylofilcon A.

In some embodiments of any one of the embodiments described herein, thehydrogel comprises a polymer consisting of poly(2-hydroxyethylmethacrylate) and/or a silicone. In some embodiments, the polymercomprises a silicone. Such polymers may optionally comprise smallamounts of additional monomers (e.g., cross-linking monomers)copolymerized with the 2-hydroxyethyl methacrylate or silicone monomer.For example, 2-hydroxyethyl methacrylate may optionally be copolymerizedwith vinyl pyrrolidone, methyl methacrylate, methacrylic acid (ananionic monomer), ethylene glycol dimethacrylate (a cross-linkingmonomer) and/or 3-(ethyldimethyl-ammonium)propyl methacrylamide (acationic monomer) in a contact lens hydrogel.

Physiological Surfaces:

In some embodiments of any of the embodiments described herein relatingto a method and/or lubrication composition for reducing a frictioncoefficient of a surface, the surface is a physiological surface, and acarrier used with the liposomes (e.g., in a lubricant compositionaccording to any of the respective embodiments described herein) is aphysiologically acceptable carrier.

In some embodiments, a surface for which a friction coefficient isreduced according any of the respective embodiments described herein isan articular surface of a synovial joint.

In some embodiments, the method of reducing a friction coefficient of asurface is for use in the treatment of a synovial joint disorderassociated with an increased friction coefficient of an articularsurface in the synovial joint.

In some embodiments of any of the embodiments described herein relatingto a liposome, the liposome is for use in the treatment of a synovialjoint disorder associated with an increased friction coefficient of anarticular surface in the synovial joint.

According to another aspect of embodiments described herein, there isprovided a use of a liposome according to any of the respectiveembodiments described herein in the manufacture of a medicament fortreating a synovial joint disorder associated with an increased frictioncoefficient of an articular surface in the synovial joint.

Examples of synovial joint disorders associated with an increasedfriction coefficient of an articular surface, and treatable according toembodiments of various aspects of the invention, include, withoutlimitation, arthritis, traumatic joint injury, locked joint (also knownin the art as joint locking), and joint injury associated with surgery.

In some embodiments, the arthritis is osteoarthritis, rheumatoidarthritis and/or psoriatic arthritis.

In some embodiments, the locked joint is associated with osteochondritisdissecans and/or synovial osteochondromatosis.

The joint injury associated with surgery described herein may optionallybe associated with surgery which directly inflicts damage on anarticular surface (e.g., by incision), and/or surgery which damages anarticular surface only indirectly. For example, surgery which repairs orotherwise affects tissue in the vicinity of the joint (e.g., ligamentsand/or menisci) may be associated with joint injury due to alteredmechanics in the joint.

The traumatic joint injury described herein may optionally be injurycaused directly by trauma (e.g., inflicted at the time of the trauma)and/or injury caused by previous trauma (e.g., a post-traumatic injurywhich develops sometime after the trauma).

The liposomes (and optionally also a water-soluble polymer describedherein) may optionally be administered as part of a composition (e.g.,solution) that comprises a physiologically acceptable carrier, forexample an aqueous carrier which is a physiologically acceptablecarrier.

Herein throughout, the term “physiologically acceptable carrier” refersto a carrier or a diluent that does not cause significant irritation toa subject upon administration in the intended manner, and does notabrogate the activity and properties of the liposomes in the composition(e.g., their ability to reduce a friction coefficient of a surface, asdescribed herein in any one of the respective embodiments). Examples,without limitations, of carriers are: propylene glycol, saline,emulsions and mixtures of organic solvents with water, as well as solid(e.g., powdered) and gaseous carriers.

Techniques for formulation and administration of compounds may be foundin “Remington's Pharmaceutical Sciences” Mack Publishing Co., Easton,Pa., latest edition, which is incorporated herein by reference.

Solutions according to any one of the embodiments of the presentinvention may be manufactured by processes well known in the art, e.g.,by means of conventional mixing or dissolving processes.

Solutions for use in accordance with the present invention thus may beformulated in conventional manner using one or more physiologicallyacceptable carriers, which facilitate processing of the liposomes (andoptionally also a water-soluble polymer described herein) intopreparations which can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen.

For injection, the liposomes described herein (optionally with awater-soluble polymer described herein) may be formulated in aqueoussolutions, preferably in physiologically compatible buffers such asHank's solution, Ringer's solution, histidine buffer, or physiologicalsaline buffer with or without organic solvents such as propylene glycol,polyethylene glycol.

The liposomes described herein (optionally with a water-soluble polymerdescribed herein) may be formulated for parenteral administration, e.g.,by bolus injection or continuous infusion. Formulations for injectionmay be presented in unit dosage form, e.g., in ampoules or in multi-dosecontainers with optionally, an added preservative. The compositions maybe suspensions, solutions or emulsions in oily or aqueous vehicles, andmay contain formulatory agents such as suspending, stabilizing and/ordispersing agents.

The liposomes described herein (optionally with a water-soluble polymerdescribed herein) may be formulated as an aqueous solution per se.Additionally, the solution may be in the form of a suspension and/oremulsions (e.g., the aqueous phase of a suspension or water-in-oil,oil-in-water or water-in-oil-in-oil emulsion), for example, in order toincrease the viscosity of the formulation. Aqueous injection suspensionsmay contain substances, which increase the viscosity of the suspension,such as sodium carboxymethyl cellulose, sorbitol or dextran. Optionally,the suspension may also contain suitable stabilizers or agents, whichincrease the solubility of the liposomes described herein (and/or theoptional water-soluble polymer described herein), for example, to allowfor the preparation of highly concentrated solutions.

In some embodiments, the liposomes described herein (optionally with awater-soluble polymer described herein) may be in powder form forconstitution with a suitable vehicle, e.g., sterile, pyrogen-free water,before use.

The solutions may be formulated wherein the liposomes are contained inan amount effective to achieve the intended purpose, for example, anamount effective to prevent, alleviate or ameliorate symptoms of adisorder in the subject being treated.

The dosage may vary depending upon the dosage form employed, the routeof administration utilized, and the location of administration (e.g.,the volume and/or surface of the region contacted with the liposomes).

The amount of a composition to be administered will, of course, bedependent on the subject being treated, the severity of the affliction,the manner of administration, the judgment of the prescribing physician,etc.

Compositions (e.g., solutions) according to embodiments of the presentinvention may, if desired, be presented in a pack or dispenser device,such as an FDA (the U.S. Food and Drug Administration) approved kit,which may contain one or more unit dosage forms containing the activeingredient(s) (e.g., liposomes described herein). The pack may, forexample, comprise metal or plastic foil, such as, but not limited to ablister pack. The pack or dispenser device may be accompanied byinstructions for administration. The pack or dispenser may also beaccompanied by a notice associated with the container in a formprescribed by a governmental agency regulating the manufacture, use orsale of pharmaceuticals, which notice is reflective of approval by theagency of the form of the compositions for human or veterinaryadministration. Such notice, for example, may be of labeling approved bythe U.S. Food and Drug Administration for prescription drugs or of anapproved product insert. Compositions comprising liposomes (optionallywith a water-soluble polymer described herein), as described herein inany one of the respective embodiments, formulated in a physiologicallyacceptable carrier may also be prepared, placed in an appropriatecontainer, and labeled for treatment of an indicated condition ordiagnosis, as is detailed herein.

Inhibition of Biofilm Formation and Biofouling:

Liposomes and bilayers described herein may optionally be useful forinhibiting adhesion, biofouling and/or biofilm formation on a surface,for example, a surface coated by a bilayer described herein, and/orcontacted with liposomes described herein.

According to an aspect of some embodiments of the invention, there isprovided a method of inhibiting adsorption of a biofouling-promotingagent on a surface of a substrate. The method, according to someembodiments of the present invention, is effected by contacting thesubstrate with a composition which comprises liposomes according to anyof the respective embodiments described herein.

The term “biofouling-promoting agent”, as used herein throughout, refersto an agent whose presence facilitates formation and/or participates information of a biofilm (as defined herein) on a substrate surface. Anagent is considered to facilitate formation of a biofilm on a substratesurface when a presence of the agent enhances formation of a biofilm ona substrate surface as compared to formation of a biofilm on the samesubstrate surface in an absence of the agent. An agent is considered toparticipate in formation of a biofilm on a substrate surface when thebiofilm formed on the surface comprises the agent as a portion of thebiofilm.

In some embodiments of any of the embodiments described herein, an agentis identified as a biofouling-promoting agent by comparing growth (e.g.,over the course of 1, 2, 3, 4, 5, 6 or 7 days) of a biofilm (e.g., P.aeruginosa) on the surface in the presence of an aqueous liquid (e.g.,water or broth, optionally at 37° C.) and the agent, to growth of abiofilm (under the same conditions) on the surface in the presence ofthe same aqueous liquid (e.g., water or broth) without the agent. Theagent is optionally mixed within the aqueous liquid, or alternatively,adsorbed onto the surface prior to exposure of the surface to theaqueous liquid. The growth of the biofilm is considered as the biofilmload at the end of the growth period (e.g., 1, 2, 3, 4, 5, 6 or 7 days)minus the initial biofilm load. Optionally, the measurement is performedsuch that the initial biofilm is substantially zero (e.g., absent or atleast undetectable), for example, the microorganism is in a planktonicform, such that growth of the biofilm is considered as the biofilm loadat the end of the growth period. In some embodiments of any of theembodiments described herein, biofilm load is defined as an area of thebiofilm.

In some embodiments of any of the embodiments described herein, biofilmload is defined as a mass and/or volume of the biofilm.

In some embodiments of any of the embodiments described herein, biofilmload is defined as a number of cells in the biofilm.

The biofilm load may optionally be determined using any technique knownin the art for detecting and quantifying an amount of cells and/ormicroorganisms in a biofilm.

In some of these embodiments, an agent is considered abiofouling-promoting agent if the growth of a biofilm in its presence isat least 10% higher than growth of a biofilm in the absence of theagent.

In some of these embodiments, an agent is considered abiofouling-promoting agent if the growth of a biofilm in its presence isat least 20% higher than growth of a biofilm in the absence of theagent.

In some of these embodiments, an agent is considered abiofouling-promoting agent if the growth of a biofilm in its presence isat least 50% higher than growth of a biofilm in the absence of theagent.

In some of these embodiments, an agent is considered abiofouling-promoting agent if the growth of a biofilm in its presence isat least 100% higher than (i.e., two-fold) the growth of a biofilm inthe absence of the agent.

Examples of biofouling-promoting agents include, without limitation, abiofouling-promoting protein and a biofouling-promoting polysaccharide,that is, any protein or polysaccharide which is a biofouling-promotingagent as defined herein.

In some embodiments of any of the embodiments described herein, thebiofouling-promoting agent is a protein.

In some embodiments of any of the embodiments described herein, themethod is considered as being capable of inhibiting adsorption of abiofouling-promoting agent when the method is capable of inhibitingadsorption of a selected biofouling-promoting agent (e.g., the selectedagent is considered representative of biofouling-promoting agents ingeneral). In some embodiments, the selected biofouling-promoting agentis a protein. In some embodiments, the selected protein is an antibodywhich does not exhibit any specific affinity to the substrate (e.g., ananti-IgG antibody, as exemplified herein).

The term “biofilm”, as used herein throughout, refers to an aggregate ofliving cells which are stuck to each other and/or immobilized onto asurface as colonies. The cells are frequently embedded within aself-secreted matrix of extracellular polymeric substance (EPS), alsoreferred to as “slime”, which is a polymeric sticky mixture of nucleicacids, proteins and polysaccharides.

In the context of the present embodiments, the living cells forming abiofilm can be cells of a unicellular microorganism, includingprokaryotes (e.g., bacteria, archaeal microorganisms) and eukaryotessuch as fungi and protists (e.g., algae, euglena, protozoa,dinoflagellates, apicomplexa, trypanosomes, amoebae) and the like; orcells of multicellular organisms, in which case the biofilm can beregarded as a colony of cells (as in the case of the unicellularorganisms) or as a lower form of a tissue.

According to some embodiments of any of the embodiments of the presentinvention, the cells are of microorganism origins, and the biofilm is abiofilm of microorganisms, such as bacteria, archaeal microorganisms,protists and fungi. The cells of a microorganism growing in a biofilmare typically physiologically distinct from cells in the “planktonic”form of the same organism, which by contrast, are single cells that mayfloat or swim in a liquid medium.

The substrate may be any substrate described herein, and encompasses anysurface, structure, product or material which can support, harbor orpromote the growth of a microorganism. The substrate is optionally aportion of an object (e.g., an article of manufacture) which cansupport, harbor or promote the growth of a microorganism. Such a portionof an object may span only a portion of an area of the object, such thata surface of the substrate represents only a portion of a surface of theobject (e.g., a portion most likely to support, harbor or promote thegrowth of a microorganism); and/or span only a portion of the thicknessof the object (e.g., along an axis perpendicular to a surface of thesubstrate and object), such that the substrate does not include theentire volume of the object lying underneath a surface of the substrate(which may represent the entire surface of the object or only a portionof the surface of the object). Non-limiting examples include the innerwalls of a storage container (e.g., a box, a can) and/or conduit (e.g.,a tubes, a pipe) for an organic product susceptible to spoilageassociated with biofouling, for example, food and/or drink (e.g., a foodcontainer, a water pipe), surfaces intended to come into contact withsuch an organic product (e.g., agricultural and/or food processingmachinery, a kitchen surface, water-purification equipment), andsurfaces exposed to moisture (e.g., bathroom walls, water systemcomponents, outer surfaces of housing exposed to rain, surfaces in thevicinity of water leakage).

In some embodiments, the substrate is a medical device or any otherdevice which is intended for contacting a living tissue, as definedherein.

In some embodiments of any of the embodiments described herein, theinhibition of adsorption described herein is for reducing adhesion ofpathogenic microorganisms (e.g., any biofilm-forming microorganismdescribed herein which is potentially pathogenic) to a medical device(e.g., any medical device described herein).

In some embodiments of any of the embodiments described herein,adsorption of the biofouling-promoting agent (any biofouling-promotingagent described herein) on the surface of the substrate subjected to amethod described herein (according to any of the respective embodiments)is reduced by at least 10% relative to adsorption on the surface of thesubstrate in the absence of the composition which comprises liposomes Insome embodiments, adsorption is reduced by at least 20%. In someembodiments, adsorption is reduced by at least 30%. In some embodiments,adsorption is reduced by at least 40%. In some embodiments, adsorptionis reduced by at least 50%. In some embodiments, adsorption is reducedby at least 60%. In some embodiments, adsorption is reduced by at least70%. In some embodiments, adsorption is reduced by at least 80%. In someembodiments, adsorption is reduced by at least 90%.

Reduction of an amount of adsorbed biofouling-promoting agent mayoptionally be determined using any technique known in the art fordetecting and quantifying an amount of agent, including, withoutlimitation, using a labeled biofouling-promoting agent (e.g., asexemplified herein in the Examples section). The reduction is optionallymeasured by contacting each of the aforementioned surfaces (e.g., for 2hours) with an aqueous solution (optionally comprising phosphate buffer,e.g., 0.1 M phosphate) of the biofouling-promoting agent (e.g., at 37°C. and/or pH 7), followed by repeated rinses to remove non-adsorbedagent (e.g., as exemplified in the examples section herein). Theconcentration of the biofouling-promoting agent in the aqueous solutionis optionally 1 μg/ml or the concentration of a saturated solution ofthe agent, whichever concentration is lower.

Herein throughout, the term “biofilm-promoting conditions” refers toconditions suitable for formation and growth of a biofilm of a cell(e.g., P. aeruginosa), for example, wherein a surface is in contact(e.g., over the course of 1, 2, 3, 4, 5, 6 or 7 days) with an aqueousliquid (e.g., water or broth, optionally at 37° C.) containing suchcells.

In some embodiments of any of the embodiments described herein, biofilmload is defined as an area of the biofilm.

In some embodiments of any of the embodiments described herein, biofilmload is defined as a mass and/or volume of the biofilm.

In some embodiments of any of the embodiments described herein, biofilmload is defined as a number of cells in the biofilm.

The biofilm load may optionally be determined using any technique knownin the art for detecting and quantifying an amount of cells and/ormicroorganisms in a biofilm.

In some embodiments of any of the embodiments described herein, the timeperiod of biofilm formation, after which biofilm load is determined, isin determined in accordance with the biofilm load, for example, the timeperiod being a time period after which a biofilm covers 100%, 50%, orany other pre-determined percentage of the area of the substrate in theabsence of inhibition of biofilm formation by contact with a compositioncomprising liposomes. For example, if a biofilm grows to cover 50% of asurface in the absence of biofilm formation inhibition, and during thesame a time period, a biofilm grows to cover 30% of a surface in thepresence of biofilm formation inhibition, then inhibition of biofilmformation may be considered to result in a reduction of 40% (i.e.,(50%-30%)/50%) in biofilm formation.

Herein, the phrase “upon contact with the agent” means that in additionto the biofilm-promoting conditions, the agent is also present (e.g., inthe aqueous liquid containing the cells).

According to an aspect of some embodiments of the invention, there isprovided a method of inhibiting biofilm formation on a surface of asubstrate (as defined herein in any embodiment and any combination ofembodiments), the method comprising contacting (as described in any ofthe respective embodiments described herein) the substrate with acomposition which comprises liposomes.

In some embodiments, “inhibiting biofilm formation” refers to theprevention of formation of a biofilm; and/or to a reduction in the rateof buildup of a biofilm; and/or to a reduction in the mass of a biofilm,the area or the volume of the biofilm, or in the number of cells formingthe biofilm.

In some embodiments of any of the embodiments described herein, theinhibition of adsorption described herein is for reducing adhesion ofpathogenic microorganisms (e.g., any biofilm-forming microorganismdescribed herein which is potentially pathogenic) to a medical device.Such a reduction may result in inhibiting biofilm formation, as definedin some embodiments herein.

In some embodiments of any of the embodiments described herein, biofilmformation on the surface of the substrate subjected to a methoddescribed herein (according to any of the respective embodiments) isreduced by at least 10% relative to biofilm formation on the surface ofthe substrate in the absence of the composition which comprisesliposomes. In some embodiments, biofilm formation is reduced by at least20%. In some embodiments, biofilm formation is reduced by at least 30%.In some embodiments, biofilm formation is reduced by at least 40%. Insome embodiments, biofilm formation is reduced by at least 50%. In someembodiments, biofilm formation is reduced by at least 60%. In someembodiments, biofilm formation is reduced by at least 70%. In someembodiments, biofilm formation is reduced by at least 80%. In someembodiments, biofilm formation is reduced by at least 90%.

The reduction in biofilm formation is optionally determined by measuringa biofilm load (in accordance with any of the respective embodimentsdescribed herein) for a biofilm of a cell (e.g., P. aeruginosa) on eachsurface after being subjected to biofouling-promoting conditions, asdefined herein (e.g., over the course of 1, 2, 3, 4, 5, 6 or 7 days, orany other time period as described herein).

Any of the embodiments described herein relating to inhibition ofbiofilm formation and/or biofouling may optionally be effected by acomposition which is essentially the same as a lubricant compositionaccording to any of the respective embodiments described herein(although optionally identified for inhibition of biofilm formationand/or biofouling rather than for lubrication).

In some of any one of the embodiments described herein which relate to ainhibition of adhesion, biofilm formation and/or biofouling, accordingto any one of the aspects described herein, the inhibition is optionallyeffected according to any of the embodiments described in Israel PatentApplication No. 234929 and/or International Patent ApplicationPCT/IL2015/050987, which are incorporated herein by reference(especially in respect to methods and compositions for inhibitingadhesion, biofilm formation and/or biofouling on a surface), with theproviso that at least a portion of the liposomes used are in accordancewith any of the embodiments described herein.

Additional Definitions

Herein, the term “hydrocarbon” describes an organic moiety thatincludes, as its basic skeleton, a chain of carbon atoms, substitutedmainly by hydrogen atoms. The hydrocarbon can be saturated ornon-saturated, be comprised of aliphatic, alicyclic or aromaticmoieties, and can optionally be substituted by one or more substituents(other than hydrogen). A substituted hydrocarbon may have one or moresubstituents, whereby each substituent group can independently be, forexample, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl,heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate,hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, oxo,cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea,thiourea, carbamate, amide, and hydrazine. The hydrocarbon can be an endgroup or a linking group, as these terms are defined herein. Thehydrocarbon moiety is optionally interrupted by one or more heteroatoms,including, without limitation, one or more oxygen, nitrogen and/orsulfur atoms. In some embodiments of any of the embodiments describedherein relating to a hydrocarbon, the hydrocarbon is not interrupted byany heteroatoms.

Preferably, the hydrocarbon moiety has 1 to 20 carbon atoms. Whenever anumerical range; e.g., “1 to 20”, is stated herein, it implies that thegroup, in this case the alkyl group, may contain 1 carbon atom, 2 carbonatoms, 3 carbon atoms, etc., up to and including 20 carbon atoms.

Herein, the term “alkyl” describes a saturated aliphatic hydrocarbon endgroup, as defined herein, including straight chain and branched chaingroups. Preferably, the alkyl group has 1 to 20 carbon atoms. Morepreferably, the alkyl is a medium size alkyl having 1 to 10 carbonatoms. Most preferably, unless otherwise indicated, the alkyl is a loweralkyl having 1 to 4 carbon atoms. The alkyl group may be substituted ornon-substituted. Substituted alkyl may have one or more substituents,whereby each substituent group can independently be, for example,cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine,halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide,sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide,and hydrazine.

The term “alkylene” describes a saturated aliphatic hydrocarbon linkinggroup, as this term is defined herein, which differs from an alkylgroup, as defined herein, only in that alkylene is a linking grouprather than an end group.

Herein, the term “alkenyl” describes an unsaturated aliphatichydrocarbon end group which comprises at least one carbon-carbon doublebond, including straight chain and branched chain groups. Preferably,the alkenyl group has 2 to 20 carbon atoms. More preferably, the alkenylis a medium size alkenyl having 2 to 10 carbon atoms. Most preferably,unless otherwise indicated, the alkenyl is a lower alkenyl having 2 to 4carbon atoms. The alkenyl group may be substituted or non-substituted.Substituted alkenyl may have one or more substituents, whereby eachsubstituent group can independently be, for example, cycloalkyl,alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate,sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy,thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy,thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.

Herein, the term “alkynyl” describes an unsaturated aliphatichydrocarbon end group which comprises at least one carbon-carbon triplebond, including straight chain and branched chain groups. Preferably,the alkynyl group has 2 to 20 carbon atoms. More preferably, the alkynylis a medium size alkynyl having 2 to 10 carbon atoms. Most preferably,unless otherwise indicated, the alkynyl is a lower alkynyl having 2 to 4carbon atoms. The alkynyl group may be substituted or non-substituted.Substituted alkynyl may have one or more substituents, whereby eachsubstituent group can independently be, for example, cycloalkyl,alkenyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate,sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy,thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy,thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.

The term “cycloalkyl” describes an all-carbon monocyclic or fused ring(i.e., rings which share an adjacent pair of carbon atoms) group whereone or more of the rings does not have a completely conjugatedpi-electron system. The cycloalkyl group may be substituted ornon-substituted. Substituted cycloalkyl may have one or moresubstituents, whereby each substituent group can independently be, forexample, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl,heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate,hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano,nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea,carbamate, amide, and hydrazine. The cycloalkyl group can be an endgroup, as this phrase is defined herein, wherein it is attached to asingle adjacent atom, or a linking group, as this phrase is definedherein, connecting two or more moieties.

The term “aryl” describes an all-carbon monocyclic or fused-ringpolycyclic (i.e., rings which share adjacent pairs of carbon atoms) endgroup (as this term is defined herein) having a completely conjugatedpi-electron system. The aryl group may be substituted ornon-substituted. Substituted aryl may have one or more substituents,whereby each substituent group can independently be, for example, alkyl,alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine,halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide,sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide,and hydrazine. Phenyl and naphthyl are representative aryl end groups.

The term “heteroaryl” describes a monocyclic or fused ring (i.e., ringswhich share an adjacent pair of atoms) group having in the ring(s) oneor more atoms, such as, for example, nitrogen, oxygen and sulfur and, inaddition, having a completely conjugated pi-electron system. Examples,without limitation, of heteroaryl groups include pyrrole, furan,thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine,quinoline, isoquinoline and purine. The heteroaryl group may besubstituted or non-substituted. Substituted heteroaryl may have one ormore substituents, whereby each substituent group can independently be,for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl,heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate,hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano,nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea,carbamate, amide, and hydrazine. The heteroaryl group can be an endgroup, as this phrase is defined herein, where it is attached to asingle adjacent atom, or a linking group, as this phrase is definedherein, connecting two or more moieties. Representative examples arepyridine, pyrrole, oxazole, indole, purine and the like.

The term “arylene” describes a monocyclic or fused-ring polycycliclinking group, as this term is defined herein, and encompasses linkinggroups which differ from an aryl or heteroaryl group, as these groupsare defined herein, only in that arylene is a linking group rather thanan end group.

The term “heteroalicyclic” describes a monocyclic or fused ring grouphaving in the ring(s) one or more atoms such as nitrogen, oxygen andsulfur. The rings may also have one or more double bonds. However, therings do not have a completely conjugated pi-electron system. Theheteroalicyclic may be substituted or non-substituted. Substitutedheteroalicyclic may have one or more substituents, whereby eachsubstituent group can independently be, for example, alkyl, alkenyl,alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine, halide,sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy,thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide,sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide,and hydrazine. The heteroalicyclic group can be an end group, as thisphrase is defined herein, where it is attached to a single adjacentatom, or a linking group, as this phrase is defined herein, connectingtwo or more moieties. Representative examples are piperidine,piperazine, tetrahydrofuran, tetrahydropyran, morpholine and the like.

As used herein, the terms “amine” and “amino” describe both a —NRxRy endgroup and a —NRx- linking group, wherein Rx and Ry are eachindependently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl orheteroalicyclic, as these terms are defined herein. When Rx or Ry isheteroaryl or heteroalicyclic, the amine nitrogen atom is bound to acarbon atom of the heteroaryl or heteroalicyclic ring. A carbon atomattached to the nitrogen atom of an amine is not substituted by ═O or═S, and in some embodiments, is not substituted by any heteroatom.

The amine group can therefore be a primary amine, where both Rx and Ryare hydrogen, a secondary amine, where Rx is hydrogen and Ry is alkyl,cycloalkyl, aryl, heteroaryl or heteroalicyclic, or a tertiary amine,where each of Rx and Ry is independently alkyl, cycloalkyl, aryl,heteroaryl or heteroalicyclic.

The terms “hydroxy” and “hydroxyl” describe a —OH group.

The term “alkoxy” describes both an —O-alkyl and an —O-cycloalkyl endgroup, or —O-alkylene or —O-cycloalkyl linking group, as defined herein.

The term “aryloxy” describes both an —O-aryl and an —O-heteroaryl endgroup, or an —O-arylene- linking group, as defined herein.

The term “thiohydroxy” describes a —SH group.

The term “thioalkoxy” describes both an —S-alkyl and an —S-cycloalkylend group, or —S-alkylene or —S-cycloalkyl linking group, as definedherein.

The term “thioaryloxy” describes both an —S-aryl and an —S-heteroarylend group, or an —S-arylene- linking group, as defined herein.

The terms “cyano” and “nitrile” describe a —C≡N group.

The term “nitro” describes an —NO₂ group.

The term “oxo” describes a ═O group.

The term “azide” describes an —N═N⁺═N⁻ group.

The term “azo” describes an —N═N-Rx end group or —N═N═ linking group,with Rx as defined herein.

The terms “halide” and “halo” refer to fluorine, chlorine, bromine oriodine.

The term “phosphate” refers to a —O—P(═O)(ORx)-OR_(Y) end group, or to a—O—P(═O)(ORx)-O— linking group, where Rx and R_(Y) are as definedherein.

The terms “phosphonyl” and “phosphonate” refer to an —P(═O)(ORx)-OR_(Y)end group, or to a —P(═O)(ORx)-O— linking group, where Rx and R_(Y) areas defined herein.

The term “phosphinyl” refers to a —PRxR_(Y) group, where Rx and R_(Y)are as defined hereinabove.

The term “sulfoxide” or “sulfinyl” describes a —S(═O)—Rx end group or—S(═O)— linking group, where Rx is as defined herein.

The terms “sulfonate” and “sulfonyl” describe a —S(═O)₂-Rx end group or—S(═O)₂— linking group, where Rx is as defined herein.

The terms “sulfonamide” and “sulfonamido”, as used herein, encompassboth S-sulfonamide and N-sulfonamide end groups, and a —S(═O)₂-NRx-linking group.

The term “S-sulfonamide” describes a —S(═O)₂-NRxR_(Y) end group, with Rxand R_(Y) as defined herein.

The term “N-sulfonamide” describes an RxS(═O)₂—NR_(Y)— end group, whereRx and R_(Y) are as defined herein.

The term “carbonyl” as used herein, describes a —C(═O)—Rx end group or—C(═O)— linking group, with Rx as defined herein.

The term “acyl” as used herein, describes a —C(═O)—Rx end group, with Rxas defined herein.

The term “thiocarbonyl” as used herein, describes a —C(═S)—Rx end groupor —C(═S)— linking group, with Rx as defined herein.

The terms “carboxy” and “carboxyl”, as used herein, encompasses bothC-carboxy and O-carboxy end groups, and a —C(═O)—O—linking group.

The term “C-carboxy” describes a —C(═O)—ORx end group, where Rx is asdefined herein.

The term “O-carboxy” describes a —OC(═O)—Rx end group, where Rx is asdefined herein.

The term “urea” describes a —NRxC(═O)—NRyRw end group or —NRxC(═O)—NRy-linking group, where Rx and Ry are as defined herein and Rw is asdefined herein for Rx and Ry.

The term “thiourea” describes a —NRx-C(═S)—NRyRw end group or a—NRx-C(═S)—NRy- linking group, with Rx, Ry and Ry as defined herein.

The terms “amide” and “amido”, as used herein, encompasses both C-amideand N-amide end groups, and a —C(═O)—NRx- linking group.

The term “C-amide” describes a —C(═O)—NRxRy end group, where Rx and Ryare as defined herein.

The term “N-amide” describes a RxC(═O)—NRy- end group, where Rx and Ryare as defined herein.

The term “carbamyl” or “carbamate”, as used herein, encompassesN-carbamate and O-carbamate end groups, and a —OC(═O)—NRx- linkinggroup.

The term “N-carbamate” describes a RyOC(═O)—NRx- end group, with Rx andRy as defined herein.

The term “O-carbamate” describes an —OC(═O)—NRxRy end group, with Rx andRy as defined herein.

The term “thiocarbamyl” or “thiocarbamate”, as used herein, encompassesO-thiocarbamate, S-thiocarbamate and N-thiocarbamate end groups, and a—OC(═S)—NRx- or —SC(═O)—NRx- linking group.

The terms “O-thiocarbamate” and “O-thiocarbamyl” describe a—OC(═S)—NRxRy end group, with Rx and Ry as defined herein.

The terms “S-thiocarbamate” and “S-thiocarbamyl” describe a—SC(═O)—NRxRy end group, with Rx and Ry as defined herein.

The terms “N-thiocarbamate” and “N-thiocarbamyl” describe a RyOC(═S)NRx-or RySC(═O)NRx- end group, with Rx and Ry as defined herein.

The term “guanidine” describes a —RxNC(═N)—NRyRw end group or—RxNC(═N)—NRy- linking group, where Rx, Ry and Rw are as defined herein.

The term “hydrazine”, as used herein, describes a —NRx-NRyRw end groupor —NRx-NRy- linking group, with Rx, Ry, and Rw as defined herein.

As used herein the term “about” refers to ±10%, and optionally ±5%.

The terms “comprises”, “comprising”, “includes”, “including”, “having”and their conjugates mean “including but not limited to”.

The term “consisting of” means “including and limited to”.

The term “consisting essentially of” means that the composition, methodor structure may include additional ingredients, steps and/or parts, butonly if the additional ingredients, steps and/or parts do not materiallyalter the basic and novel characteristics of the claimed composition,method or structure.

As used herein, the singular form “a”, “an” and “the” include pluralreferences unless the context clearly dictates otherwise. For example,the term “a compound” or “at least one compound” may include a pluralityof compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention maybe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 3, 4, 5, and 6. This appliesregardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to includeany cited numeral (fractional or integral) within the indicated range.The phrases “ranging/ranges between” a first indicate number and asecond indicate number and “ranging/ranges from” a first indicate number“to” a second indicate number are used herein interchangeably and aremeant to include the first and second indicated numbers and all thefractional and integral numerals therebetween.

As used herein the term “method” refers to manners, means, techniquesand procedures for accomplishing a given task including, but not limitedto, those manners, means, techniques and procedures either known to, orreadily developed from known manners, means, techniques and proceduresby practitioners of the chemical, pharmacological, biological,biochemical and medical arts.

As used herein, the term “treating” includes abrogating, substantiallyinhibiting, slowing or reversing the progression of a condition,substantially ameliorating clinical or aesthetical symptoms of acondition or substantially preventing the appearance of clinical oraesthetical symptoms of a condition.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination or as suitable in any other describedembodiment of the invention. Certain features described in the contextof various embodiments are not to be considered essential features ofthose embodiments, unless the embodiment is inoperative without thoseelements.

Various embodiments and aspects of the present invention as delineatedhereinabove and as claimed in the claims section below find experimentalsupport in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with theabove descriptions illustrate some embodiments of the invention in anon-limiting fashion.

Materials and Methods

Materials:

2-Bromoisobutyryl bromide was obtained from Sigma-Aldrich.

Chloroform was obtained from Sigma-Aldrich.

CuBr was obtained from Sigma-Aldrich.

Dichloromethane was obtained from Sigma-Aldrich.

Dimyristoylphosphatidylcholine (DMPC) was obtained from Lipoid GmbH.

Distearoylphosphatidylethanolamine (DSPE) was obtained from Avanti PolarLipids.

Distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG, withPEG Mw of 2000 Da (DSPE-PEG 2000) or 5000 Da (DSPE-PEG 5000)) andDipalmitoylphosphatidylethanolamine-polyethylene glycol (DPPE-PEG 2000,PEG Mw of 2000 Da) were obtained from Avanti.

1,2-Distearoyl-sn-glycerol (DSG) was obtained from Santa CruzBiotechnology (Germany).

Ethanol was obtained from Bio-Lab.

Hydrogenated soy phosphatidylcholine (HSPC) was obtained from LipoidGmbH.

Methanol was obtained from Bio-Lab.

N,N,N′,N″,N″-pentamethyldiethylenetriamine (PMDETA) was obtained fromSigma-Aldrich.

O-(2-methacryloyloxyethyl)phosphorylcholine (MPC) was obtained fromBiocompatible Corporation (UK).

Phosphate buffer saline (PBS) was obtained from Sigma-Aldrich.

Triethylamine was obtained from Sigma-Aldrich.

Water was purified using a Barnstead NanoPure system to 18.2 MΩ cmresistance with total organic content levels of <ca.1 ppb.

Etafilcon A (1-Day ACUVUE®) and Narafilcon A (1-Day TruEye®) contactlenses were obtained from Johnson & Johnson, immersed in saline solutionin a blister-pack. The composition, water content and modulus of thecontact lenses (as stated by the manufacturer) are as follows. EtafilconA lenses contain 2-hydroxyethylmethacrylate (HEMA) and methacrylic acid(MA), have a water content of 58%, and a modulus of 0.3 MPa. NarafilconA lenses contain silicone, have a water content of 46%, and a modulus of0.66 MPa.

Atomic Force Microscopy (AFM) and Surface Force Balance (SFB)Measurements of Liposomes:

Liposome-covered mica surfaces were prepared by placing freshly cleavedmica in a 1 mM SUV liposome dispersion prepared with pure water or withwater at pH 5 or with 0.15 M NaNO₃, at room temperature. After overnightincubation, the surfaces were rinsed to remove excess material byplacing them in a beaker containing 300 ml pure water or 0.15 M NaNO₃for 30 minutes, using a delicate shaking motion. Surfaces prepared forSFB always passed the air-water interface, whereas for examination byAFM, the surfaces did not pass an interface unless the effect of passingan interface was being examined. In the contact lens experimentsdescribed below, the treated contact lens did pass the air-waterinterface.

Contact Lens Treatment and Measurements:

Contact lenses were removed from their container, where they had beenimmersed in PBS solution, and then rinsed with PBS. The lenses were thenimmersed for 2-5 days in the indicated solution of polymer and/orliposomes in PBS, or in PBS alone (as a control). Lipid concentrationsin the liposome solutions were 10 mM. Following immersion for 2-5 days,the lenses were in all cases thoroughly rinsed by a stream of PBS priorto measurements in a tribometer.

Friction tests were performed with a UMT model tribometer (Bruker).Contact lenses were mounted on a cornea-mimicking holder, which has atypical geometry resembling the human cornea, as shown in FIGS. 8A and8B. The contact lens was then positioned opposite a glass plate andimmersed in PBS during the measurement. The normal loads that were usedwere of 3 grams and 10 grams. The friction coefficient was calculated bydividing the measured lateral force by the applied normal force.Friction coefficient values are those of kinetic friction, which isrelated to the forces in the system that are measured when there is asliding motion of the contact lens on the opposing glass surface.Parameters were as follows: sliding velocity 1 mm/second, amplitudes of1-1.5 mm, frequency 1 Hz, and dwell time of 5 seconds prior toinitiation of motion. Experiments were conducted at 37±1° C., and thefriction values were taken as an average over 300 cycles for each datapoint.

The mean pressure P over the contact area A was evaluated as P=F_(N)/A,where F_(N) is the applied normal load and, from Hertzian contactmechanics [Johnson, Contact Mechanics 2004, London: Cambridge UniversityPress], A=π(R F_(N)/k)^(2/3), where R is the radius of the rigidcornea-mimicking holder and k is the Young modulus of the contact lens,as provided by the manufacturer.

Example 1 Phospholipid with Polymerized Phosphocholine Derivative

The overall preparation of a phospholipid with a polymerizedphosphocholine derivative, from the phospholipid DSPE(distearoylphosphatidylethanolamine) and the phosphocholine derivativeMPC (O-(2-methacryloyloxyethyl)phosphorylcholine), is depictedschematically in Scheme 1 below. The obtained polymeric product isreferred to herein as DSPE-PMPC(distearoylphosphatidylethanolamine-substitutedpoly(O-(2-methacryloyloxyethyl)phosphorylcholine)). DSPE-PMPC may beregarded as a polymeric phosphatidylcholine analog in view of thesimilarity between the structure of each MPC unit and the structure of aphosphocholine head group in phosphatidylcholine.

Synthesis of DSPE-Br Initiator:

Triethylamine (0.3 ml, 2.35 mmol) was added to 30 ml of dried chloroformcontaining 0.88 gram (1.17 mmol) of distearoylphosphatidylethanolamine(DSPE), and the mixture was stirred at room temperature for 0.5 hour.2-bromoisobutyryl bromide (0.115 ml, 1.17 mmol) was then injected intothe solution. The mixture was then stirred overnight at 40° C. Thesolution was washed with water three times, and a white power wasobtained by removing the solvent using a rotary evaporator.

Synthesis of DSPE-PMPC by Atom-Transfer Radical Polymerization (ATRP):

DSPE-Br (87 mg, 0.1 mmol) was dissolved in 3 ml of dichloromethane, and740 mg (2.5 mmol) of MPC (O-(2-methacryloyloxyethyl)phosphorylcholine)was dissolved in 9 ml of ethanol. DSPE-Br, MPC and CuBr (14 mg, 0.1mmol) were placed in a Schlenk flask with a magnetic stirrer bar. Theflask was degassed by nitrogen for 30 minutes. 40 μl (0.2 mmol) ofPMDETA (N,N,N′,N″,N″-pentamethyldiethylenetriamine) was then injectedquickly. The flask was stirred for 16 hours at 60° C. Then the solutionwas dialyzed against ethanol (in order to remove catalyst and ligand)and against water (in order to remove unreacted monomer), for 48 hourseach (Mw cutoff of 3500 Da). The polymer DSPE-PMPC was obtained afterfreeze-drying.

As shown in FIG. 1, the amount of carbon atoms attached to a phosphategroup was similar to the amount of carbon atoms in a stearoyl alkylchain, as determined by ¹H-NMR spectroscopy. This result indicates thatthe degree of polymerization of DSPE-PMPC was about 17 MPC units(corresponding to a Mw of about 5 kDa) per DSPE moiety.

Example 2 Stabilized Liposomes Containing DSPE-PMPC

To prepare liposomes, 178 mg of hydrogenated soy phosphatidylcholine(HSPC) and 20 mg of DSPE-PMPC (prepared as described in Example 1) weredissolved in methanol and chloroform (2 ml, 1:1 v/v). The organicsolvent was then dried by nitrogen overnight. Multilamellar vesicles(MLVs) of modified HSPC were prepared in pure water or in 0.15 M NaNO₃by sonication for 15 minutes at 70° C., and then downsized to form smallunilamellar vesicles (SUVs) of about 70 nm in diameter, at aconcentration of 15 mM (by phospholipid concentration), by stepwiseextrusion through polycarbonate membranes, starting with 400 nm (5cycles), 100 nm (6 cycles) and ending with 50 nm (8 cycles). MLVs ofHSPC alone were prepared in pure water using standard techniques anddownsized to form SUVs by extrusion.

The stability of liposomes was checked by dynamic light scattering (DLS)measurements.

As shown in FIG. 2, after one month at 4° C., HSPC SUVs formedaggregates (characterized by a peak at around 1000 nm), whereas noaggregation of HSPC SUVs with DSPE-PMPC was observed, indicating thatthe DSPE-PMPC stabilized the liposomes.

This result suggests that the PMPC moieties in DSPE-PMPC forms tailswhich extend from the liposomes into the surrounding environment,thereby forming a steric barrier to aggregation and fusion.

Surfaces with adsorbed DSPE-PMPC-stabilized liposomes were characterizedby atomic force microscopy, according to procedures described in theMaterials and Methods section hereinabove.

As shown in FIG. 3, no individual liposomes were observed by atomicforce microscopy on a mica surface that was coated withDSPE-PMPC-stabilized liposomes incubated in water (pH 5.8).

As shown in FIG. 4, closely packed and stably adsorbedDSPE-PMPC-stabilized liposomes were observed by atomic force microscopyon a mica surface incubated in water at pH 5.

These results indicate that lowering the pH to 5 neutralizes enoughnegative charges in the liposomes to allow adsorption of the liposomesonto the negatively charged mica surface.

The properties of DSPE-PMPC-stabilized HSPC liposomes were examined inwater and in salt (0.15 M NaNO₃) solutions, and compared to those ofDSPE-PEG 5000-stabilized HSPC liposomes. Friction forces were evaluatedunder various loads, and the sliding friction coefficient (μ) wasevaluated simply by dividing the friction force by the load.

As shown in FIGS. 5A and 6A, the normal forces between the surfaces (Fn)increases as the separation distance (D) between surfaces decreases, anda sharp increase of Fn is observed at close separation. The finalhigh-pressure separation was 20 nm in water (FIG. 5A) and 12 nm in saltsolution (FIG. 6A).

As shown in FIG. 5B, DSPE-PEG-stabilized HSPC layers could withstandpressures up to 80 atm with a friction coefficient of about 0.01 inwater, whereas DSPE-PMPC-stabilized HSPC could sustain pressure at least100 atm with a friction coefficient of about 0.002-0.008 in water.

These results indicate that DSPE-PMPC-stabilized HSPC exhibits excellentlubrication properties, in terms of both reduced friction coefficientand endurance under high pressure.

These results further indicate that DSPE-PMPC-stabilized lipid layersexhibit superior lubrication under high pressures (e.g., 80 atm or more)as compared with PEG-stabilized lipid layers.

In an earlier study [Goldberg et al., Adv Materials 2011, 23:3517-3521],PEG-stabilized liposomes exhibited considerably poorer lubrication inpure water. Without being bound by any particular theory, thedifferences between the results presented herein for PEG-stabilizedliposomes and the aforementioned results of Goldberg et al. may be dueto fact that the results reported in Goldberg et al. were obtained usingliposomes/PEG adsorbed onto a solid mica surface under pressures of upto 10 atm, whereas results described herein are obtained with softhydrogel surfaces at pressures of only about 0.1 atm; and/or due to thefact that the PEG-terminated lipids used in Goldberg et al. had only C14chains, whereas the PEG-terminated lipid used in the present results(DSPE-PEG) contains C18 chains.

As shown in FIG. 6B, in a salt solution, DSPE-PEG-stabilized HSPC layersexhibited friction coefficients of ≈0.01 and could withstand pressuresonly up to about 20-40 atm, whereas DSPE-PMPC-stabilized HSPC exhibitedfriction coefficients of about 0.001 or less and could sustain pressuresof at least 100 atm.

These results indicate that the presence of salt, which representsconditions similar to physiological conditions, reduces the lubricationefficacy of PEG-stabilized lipids, in terms of both the ability to lowerfriction coefficients and endurance under high pressure, whereas thelubrication efficacy of DSPE-PMPC-stabilized lipids is maintained in thepresence of salt, and in comparison to DSPE-PEG, lubrication efficacy ofDSPE-PMPC-stabilized lipids is better in the presence of salt.

Furthermore, as shown in FIG. 7, 20 mg/ml of DSPE-PMPC incubated in purewater (at pH 5) for one day formed micelles, indicating that DSPE-PMPCcan form a stable dispersion even in the absence of another lipid.

Without being bound by any particular theory, it is believed that suchmicelles expose highly-hydrated phosphocholine groups, and can providevery good boundary lubrication (e.g., between compressed mica surfaces)up to at least moderate pressures (e.g., 25 atm).

Example 3 Effect of Stabilized Liposomes on Contact Lens FrictionCoefficients

In order to prepare stabilized liposomes, DSPE-PMPC (prepared asdescribed in Example 1, except that the Mw of the PMPC moiety was about2 kDa) and HSPC at molar ratio of 2:98 were dissolved in organic solventand left overnight under nitrogen to form a dry film. Multilamellarvesicles (MLV) were then prepared by hydrating the lipids at least 5° C.above the lipid melting point, followed by sonication, in PBS. The sameprocedure was also applied on the following mixtures: DPPE-PEG 2000 andDMPC, DSPE-PEG 2000 and HSPC, both at molar ratio of 2:98, to form MLVs.The organic solvents that were used are chloroform for the HSPC andDMPC, methanol for the DSPE-PMPC and ethanol for the DPPE-PEG 2000 andDSPE-PEG 2000.

MLVs were downsized to form small unilamellar vesicles (SUV), ˜100 nm indiameter, by stepwise extrusion through polycarbonate membranes startingwith a 400-nm and ending with 50-nm pore-size membrane, using a Lipex 10ml extruder system (Northern Lipids, Canada). Dynamic light scatteringmeasurements (DLS) revealed a diameter of ˜100 nm for all liposomes.

Friction coefficients were measured for etafilcon A and narafilcon Acontact lenses in a PBS environment following 2 day immersion in PBSsolutions with or without liposomes, according to the proceduresdescribed in the Materials and Methods section hereinabove.

As shown in FIGS. 9 and 10, the liposomes solutions with eitherDSPE-PMPC or PEGylated lipids (DPPE-PEG 2000 or DSPE-PEG 2000) eachreduced the friction coefficient of the contact lenses to a considerabledegree, under each of the tested loads.

Example 4 Glycerolipid with Polymerized Phosphocholine Derivative

A lipid with polymerized phosphocholine derivative was preparedaccording to procedures similar to those described in Example 1, exceptthat a glycerolipid without a phosphate group,1,2-distearoyl-sn-glycerol (DSG), was used instead of DSPE. The overallpreparation of a lipid (referred to herein as DSG-PMPC) with apolymerized phosphocholine derivative, from DSG and the phosphocholinederivative MPC (O-(2-methacryloyloxyethyl)phosphorylcholine), isdepicted schematically in Scheme 2 below.

Synthesis of DSG-Br Initiator:

Triethylamine (0.3 ml, 2.35 mmol) was added to 30 ml of dried chloroformcontaining 0.92 gram (1.17 mmol) of 1,2-distearoyl-sn-glycerol (DSG),and the mixture was stirred at room temperature for 0.5 hour.2-bromoisobutyryl bromide (0.115 ml, 1.17 mmol) was then injected intothe solution. The mixture was then stirred overnight at 40° C. Thesolution was washed with water three times, and a white power wasobtained by removing the solvent using a rotary evaporator.

Synthesis of DSPE-PMPC by Atom-Transfer Radical Polymerization (ATRP):

DSG-Br (94 mg, 0.1 mmol) was dissolved in 3 ml of dichloromethane, and740 mg (2.5 mmol) of MPC (O-(2-methacryloyloxyethyl)phosphorylcholine)was dissolved in 9 ml of ethanol. DSG-Br, MPC and CuBr (14 mg, 0.1 mmol)were placed in a Schlenk flask with a magnetic stirrer bar. The flaskwas degassed by nitrogen for 30 minutes. 40 μl (0.2 mmol) of PMDETA(N,N,N′,N″,N″-pentamethyldiethylenetriamine) was then injected quickly.The flask was stirred for 16 hours at 60° C. Then the solution wasdialyzed against ethanol (in order to remove catalyst and ligand) andagainst water (in order to remove unreacted monomer) for 48 hours each(Mw cutoff of 3500 Da). The polymer DSG-PMPC was obtained afterfreeze-drying.

¹H-NMR spectroscopy indicated that the degree of polymerization ofDSG-PMPC was about 16 MPC units (corresponding to a Mw of about 5 kDa)per DSG moiety (data not shown).

As shown in FIG. 11, the net charge of the lipid derivative was zero, asdetermined by zeta potential measurements.

Example 5 Stabilized Liposomes Containing DSG-PMPC

To prepare liposomes, 178 mg of hydrogenated soy phosphatidylcholine(HSPC) and 22 mg of DSG-PMPC (prepared as described in Example 4) weredissolved in methanol and chloroform (2 ml, 1:1 v/v). The organicsolvent was then dried by nitrogen overnight. Multilamellar vesicles(MLVs) of modified HSPC were prepared in pure water by sonication for 15minutes at 65° C., and then downsized to form small unilamellar vesicles(SUVs) of about 70 nm in diameter, at a concentration of 30 mM (byphospholipid concentration), by stepwise extrusion through polycarbonatemembranes, starting with 400 nm (5 cycles), 100 nm (6 cycles) and endingwith 50 nm (12 cycles).

The stability of liposomes was checked by dynamic light scattering (DLS)measurements. After one month at 4° C., no aggregation of HSPC SUVs withDSG-PMPC was observed, indicating that the DSG-PMPC stabilized theliposomes. This result suggests that the PMPC moieties in DSG-PMPC formstails which extend from the liposomes into the surrounding environment,thereby forming a steric barrier to aggregation and fusion.

Surfaces with adsorbed DSG-PMPC-stabilized liposomes were characterizedby atomic force microscopy, according to procedures described in theMaterials and Methods section hereinabove.

As shown in FIG. 12, stably adsorbed DSG-PMPC-stabilized liposomes wereobserved by atomic force microscopy on a mica surface incubated in water(pH 5.8).

In contrast, as shown in FIG. 3, negatively charged DSPE-PMPC-stabilizedHSPC liposomes did not adsorb on mica in water at pH 5.8.

These results indicate that neutrally charged DSG-PMPC allows foradsorption to surfaces which repel negatively charged compounds such asDSPE-PMPC.

As shown in FIG. 13, DSG-PMPC-stabilized HSPC could sustain pressures ofat least 100 atm with a friction coefficient of about 0.003-0.006 inwater.

These results indicate that DSG-PMPC-stabilized HSPC exhibits excellentlubrication properties.

Although the invention has been described in conjunction with specificembodiments thereof, it is evident that many alternatives, modificationsand variations will be apparent to those skilled in the art.Accordingly, it is intended to embrace all such alternatives,modifications and variations that fall within the spirit and broad scopeof the appended claims.

All publications, patents and patent applications mentioned in thisspecification are herein incorporated in their entirety by referenceinto the specification, to the same extent as if each individualpublication, patent or patent application was specifically andindividually indicated to be incorporated herein by reference. Inaddition, citation or identification of any reference in thisapplication shall not be construed as an admission that such referenceis available as prior art to the present invention. To the extent thatsection headings are used, they should not be construed as necessarilylimiting.

What is claimed is:
 1. A polymeric compound having the general formulaI:

wherein: m is zero or a positive integer; n is an integer which is atleast 1, wherein when X does not comprise a phosphate group, n is atleast 2; X is a lipid moiety; Y is a backbone unit which forms apolymeric backbone; L is absent or is a linking moiety; and Z has thegeneral formula II:

wherein: A is a substituted or unsubstituted hydrocarbon; B is an oxygenatom or is absent; and R₁-R₃ are each independently selected from thegroup consisting of hydrogen, alkyl, cycloalkyl, heteroalicyclic, aryland heteroaryl, and wherein: a) said lipid is selected from the groupconsisting of a fatty acid, a monoglyceride, a glycerophospholipid, anda sphingolipid, and/or b) X has the general formula III:

wherein: W₁ and W₂ are each independently selected from the groupconsisting of hydrogen, alkyl, alkenyl, alkynyl and acyl, wherein atleast one of W₁ and W₂ is not hydrogen; J is —P(═O)(OH)—O— or absent; Kis a substituted or unsubstituted hydrocarbon from 1 to 10 carbon atomsin length; M is a linking group selected from the group consisting of—O—, —S—, amino, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl,carbonyl, thiocarbonyl, urea, thiourea, carbamyl, thiocarbamyl, amido,carboxy, and sulfonamide, or absent; and Q is a substituted orunsubstituted hydrocarbon from 1 to 10 carbon atoms in length, orabsent, wherein when M is absent, Q is also absent.
 2. The polymericcompound of claim 1, wherein Y is a substituted or unsubstitutedalkylene unit.
 3. The polymeric compound of claim 2, wherein Y has theformula —CR₄R₅—CR₆D-, wherein: when Y is a backbone unit which is notattached to said L or said Z, D is R₇; and when Y is a backbone unitwhich is attached to said L or said Z, D is a covalent bond or a linkinggroup attaching Y to said L or said Z, said linking group being selectedfrom the group consisting of —O—, —S—, alkylene, arylene, sulfinyl,sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl,urea, thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, and amino; andR₄-R₇ are each independently selected from the group consisting ofhydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl,heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy,thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, azide, azo,phosphate, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea,thiourea, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, sulfonamido, and amino.
 4. Thepolymeric compound of claim 1, wherein A is a substituted orunsubstituted hydrocarbon from 1 to 4 carbon atoms in length.
 5. Thepolymeric compound of claim 1, wherein n is at least
 3. 6. The polymericcompound of claim 1, wherein at least a portion of said Y, said L and/orsaid Z comprise at least one targeting moiety.
 7. The polymeric compoundof claim 1, wherein said lipid is selected from the group consisting ofa fatty acid, a monoglyceride, a glycerophospholipid, and asphingolipid.
 8. The polymeric compound of claim 1, wherein X has saidgeneral formula III.
 9. The polymeric compound of claim 8, wherein atleast one of W₁ and W₂ is alkyl, alkenyl, alkynyl or acyl, being from 10to 30 carbon atoms in length.
 10. A lipid bilayer comprising at leastone bilayer-forming lipid and the polymeric compound of claim
 1. 11. Aliposome comprising at least one lipid bilayer according to claim 10.12. The liposome of claim 11, further comprising at least one functionalmoiety or agent, bound to a surface of the liposome and/or within alipid bilayer and/or core of the liposome.
 13. A lubricant compositioncomprising liposomes according to claim 11 and a carrier.
 14. Thelubricant composition of claim 13, being for lubricating a physiologicalsurface, wherein said carrier is a physiologically acceptable carrier.15. A method of reducing a friction coefficient of a surface, the methodcomprising contacting the surface with liposomes according to claim 11.16. The method of claim 15, further comprising contacting the surfacewith a water-soluble polymer.
 17. The method of claim 15, wherein saidsurface is a physiological surface, the method being effected bycontacting the surface with a composition comprising said liposomes anda physiologically acceptable carrier.
 18. A method of treating asynovial joint disorder associated with an increased frictioncoefficient of an articular surface in the synovial joint in a subjectin need thereof, the method comprising administering to the subject theliposome of claim
 11. 19. A method of inhibiting biofilm formation on asurface of a substrate, the method comprising contacting the substratewith a composition which comprises liposomes according to claim
 11. 20.An article of manufacture comprising a composition-of-matter, thecomposition-of-matter comprising a substrate coated, on at least aportion of a surface thereof, by a lipid bilayer according to claim 10.